Applied Biotechnology - Science topic

The use of biotechnology to carry out species protection does provide an innovative way to prevent species extinction. CRISPR-Cas9 and other technologies enhance the adaptability of animals and plants to climate change, disease and pollution, and in vitro breeding technology builds a protective barrier for species survival at the technical level by assisting the breeding of endangered species. However, the actual effectiveness of such technologies is highly dependent on political policy support (e.g., biosafety regulatory frameworks, conservation funding) and social values (e.g., public acceptance of gene-editing technology), and more fundamentally on the need to simultaneously promote systemic adjustment of human activities - only by effectively curbing destructive practices such as deforestation and overexploitation. To really eliminate the primary drivers of extinction. At the same time, ethical disputes always accompany the process of technology application: whether artificial intervention in animal genomes deviates from the natural conservation philosophy of "let nature take its course", the ecological niche conflict that may arise from the revival of extinct species, and the risk of technological abuse need to be included in the system of scientific ethical review and social co-governance. Therefore, the successful practice of species conservation is essentially the result of the synergistic effect of technical means, policy regulation, social consensus and ethical considerations.

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Dear Researchers, Scientists, and Friends,

The global water crisis demands urgent, innovative, and sustainable solutions. Biotechnology offers promising pathways for safeguarding and regenerating drinking water resources while addressing global water shortages. Through genetically engineered microorganisms and bio-based processes, biotechnology enhances water purification systems by breaking down harmful pollutants, treating wastewater, and even restoring contaminated aquifers.

One critical contribution lies in desalination and purification. Bio-desalination, using salt-tolerant microbial strains and biomimetic membranes, can reduce energy consumption compared to traditional thermal or reverse osmosis methods. Similarly, biofiltration systems, relying on microbial consortia, can purify water more efficiently while minimizing chemical usage. Biotechnology also supports real-time water quality monitoring through biosensors, enabling rapid detection of toxins and pathogens.

However, implementation challenges remain—most notably high costs, energy demands, and potential ecological impacts. Hence, interdisciplinary collaboration is vital, integrating environmental engineering, economics, and sustainability sciences to design scalable, eco-friendly systems.

In this context, the principles of the green economy and circular economy become instrumental. Biotechnology aligns naturally with these models by transforming waste into resources, promoting water reuse, and reducing reliance on finite inputs. Wastewater, once seen as a burden, can become a biotechnological asset—producing clean water, biogas, and nutrients for agriculture.

In conclusion, biotechnology is not a silver bullet, but with targeted research and cross-sector integration, it can be a cornerstone in the global shift toward sustainable water resource management and a circular water economy.

Where can I send these bacteria for chemotaxonomic characterization (FAME analysis and polar lipid)?

I don't have an answer, but this is a very impressive question!!!!!

Thank you for the accurate and interesting views. I'm actually wondering the considerations one would make, when he/she chooses a target gene and vector design. More specifically the genetic consideration in replacing or adding a gene into mammalian cell types.

Hi,

Bioinformatics is a promising career option both in India and abroad.

for creating graphic images I recommend you biorender.com you have on this website all the templates also they are for free, you can register and start creating your first graphic abstract and images. in many articles and reviews, all the graphics are made via this website.

- You've made excellent points with clear analogies - thank you! I agree with your synopsis that "Transitioning biological understanding to this next level is certainly in large part a logistical problem" and that we must strive to the "coordination of manpower and computational power resources being put towards the pursuit of understanding things at this level."

Hi

In Iran, you can order your strain to Iranian Biological Resource Center (IBRC).

Please check www.ibrc.ir

I would propose to use an experimental setting with low basal VEGF expression. However, depending on your goal you should consider to use a cell type which is involved in the process, you want to study (e.g. for vascularization -> endothelail cells, pathophysiological relevat cell growth -> tumor cells etc.).

we have contacted qiagen and they have proposed us the PRO version of the kit. Of course the results with the normal kit and the PRO are not comparable, that's why we would like to run all the samples with the same type of kit for DNA extraction. Anyway thanks for checking in your lab. if you had some of them

Best regards

Microwave (or an oven) will act fine, killing the tissue. But change paper frequently after applying it, especially with succulents.

DNA fragment to be cloned must be isolated, you do with PCR or restriction enzymes isolated from bacteria, which are able to recognize and cut specific sequences creating "Sticky" or "Blunt" DNA ends.

The plasmid is digested with restriction enzymes, opening up the vector to allow insertion of the target DNA. If the isolated DNA of interest and the plasmid or vector are digested with the same restriction enzyme, their sticky ends will be complementary. The two DNAs are then incubated with DNA ligase, an enzyme that can attach together strands of DNA with double strand breaks.

Following ligation, the recombinant DNA is placed into a host cell like E. Coli, in a process called transfection or transformation with high concentration of calcium or heat shocks.

Finally, the transfected cells are then cultured but some may not contain a plasmid with the target DNA because the transfection process is not usually 100% successful and the appropriate cultures must be selected with markers usually for antibiotic resistance. When the treated cells are plated on a petri dish of nutrient agar containing the antibiotic, only the rare transformed cells containing the antibiotic-resistance gene on the plasmid vector will survive.

Further analysis of the resulting colonies is required to confirm that cloning was successful. This may be accomplished by means of a process PCR or restriction fragment analysis, both of which need to be followed by gel electrophoresis and/or DNA sequencing. DNA sequence analysis, PCR, or restriction fragment analysis will all determine if the plasmid/vector contains the insert. Restriction fragment analysis is digestion of isolated plasmid/vector DNA with restriction enzymes. If the isolated DNA contains the target DNA, that fragment will be excised by the restriction enzyme digestion. Gel electrophoresis will separate DNA molecules based on size and charge.

I hope I was helpful Golla Ramanjaneyulu

1. C1*V1 = C2*V2

200 mg/ml * V1 = 40 ug/ml * 300 ml, or V1 = 0.06 ml (of ampiciline)

2. If you mean 75 ml is the total volume of final medium, then:

C1 * V1 = 75 ml * C1/50

V1 = 75/50 = 1.5 ml (of bacteria)

molecular virology

Hi Logan,

Since you already have interested suppliers, why not contact their customer service for quotations? Different companies have different technical support services and final delivery quality, so it’s difficult to compare, depending on whether you care more about data quality or cost.

Both RT-PCR and Qualitative PCR (Endpoint) are essentially the same PCR process. The difference comes that RT-PCR collects signal (florescent) information at the end of each amplification cycle building the amplification curve where with endpoint PCR you are limited with the signal at the end of the process. After RT-PCR is finished the product in the plate can be run on the get as endpoint PCR amplicons.

Endpoint PCR, in general, can only reliably answer if this genetic sequence determined by the primer pair is present in the sample. RT-PCR, however, can detect the difference in abundance.

For example, you have the 3 samples of transgenic mice: with the gene of interest presented as follows: +/+, +/-, and -/- if you run endpoint PCR in most cases you get positive bands in first and second samples and no band in third. You will not be able to reliably determine between +/+ and +/-. If you run the RT-PCR however on the same samples you will clearly see the difference in the amplification curve.

I.e. if we use RT-PCR and add histone primers group as our housekeeper and do the normalization the values for the example sample would be as follows: +/+ (1.0) , +/ (0.5)-, -/- (0 no amplification). Of course, since it is relative abundance we can choose any sample (exempt 0) to be our 1 (or 100 if you like). For example, if we choose our +/- to be 1 the values change to +/+ (2.0) , +/ (1.0)-, -/- (0 no amplification) representing the same results.

In most common use, however, the RT-PCR utilized to measure gene expression through extracting the total RNA from the sample, conversion of RNA to c-DNA and then amplification of c-DNA with PCR chemistry with measurement of fluorescence at each step. If amplification curves are normalized to the housekeeping genes such as GAPDH or B-actin relative abundance can be determined as fold change value.

Some scientists on the limited budget even try to use quantification with the endpoint PCR in a similar way using 4 primers in the mix for the target and housekeeping genes and then performing relative quantification of the bands on the gel. (the same way Western Blot samples are quantified). It sometimes works, but it is not a very sensitive method or with RT-PCR being cheap and available not very well accepted by the scientific community. In the past, I have seen people doing it.

In theory, you can replicate the RT-PCR curve process using standard PCR and agarose gel (i.e. without RT-PCR machine). You will need to prepare at least 6 (More is more reliable) or so samples and then run 6 amplification programs for the progressive increase of the number of cycles i.e. stop the program after 5, 10, 15, 20, 25, 30. Then run all samples on the gel and see at which cycle you get the band visible on the gel this will be your Ct value (cycle threshold). And then run the same process for your housekeeper to make sure that housekeeping gene bands will show up in the same cycle if there is a difference in housekeeping band than you to make a correction for the cycle number i.e. if one sample produce band for housekeeper earlier than other than there is a difference in loading that need to be normalized. Naturally, this method is very resource and time/effort consuming and not very reliable. I believe back in the 1980es people might have tried to do something like this but nowdays everything is automated.

In the context of viral infections, the difference between the endpoint and RT-PCR is the following: Endpoint PCR will let you know if the virus present or absent even if there is a few copies of the virus after 30 cycles PCR will produce the signal 2^29 is a huge amplification power.

If you run RT-PCR for the virus you can determine the viral load. The earlier cycle threshold will indicate more virus copies present in the initial sample and more severe infection. For example, you have 3 patients samples and one healthy control. You got Ct values: 8, 10, 30 and 0 (no signal) for control. These results will clearly indicate that patient 1 with (ct value 8 has the most severe infection) as the sample was amplified to be detectable only after 8 cycles amplification 2^7 meaning very high initial copy number, patient 2 with ct value 15 is has a mild infection and the last patient have very few copies of the virus that was amplified only after 30 cycles 2^29. If you run PCR products on the gel you might even see these differences if they are explicit. i.e. the first patient band would be brighter. (that is the meaning of qualitative PCR). However, RT-PCR allows you to distinguish very subtle changes. I don't think it is very meaningful to use any housekeeping gene when working with the virus material so you can operate directly with the ct values. There are few techniques where you can convert ct values to the viral copy numbers, this was determined in the literature for most known viruses usually by building the linear or log curve of the known copy numbers and projecting the ct values to the curve.

I hope it helps your understanding of the PCR and RT-PCR and its use with the viruses.

Herbal medicines may produce negative effects such as allergic reactions, rashes, asthma, headaches, nausea, vomiting, and diarrhoea that can range from mild to severe.

How to kill those bacteria?

Hello, there were many aspects in microbiology which need more detail study till now we have information only about 5% of total biodiversity of microorganisms. So 95% is future work!

Good luck!

On the plant surface (and in solution), paraquat is rapidly broken down photochemically.

Considering probiotics and / or prebiotics as alternatives or substitutes for antibiotics would be very compromising to affirm, since these do not act like antibiotics, the modes of action are very different. Although these ingredients provide certain benefits to human and animal health, to work properly depends on many factors, and its effects are medium and long term, while antibiotics have an immediate effect in the short term. Some probiotic strains produce certain proteins with antimicrobial effects, such as bacteriocins, among them, we have Nisin, which is used as a broad spectrum preservative in the food industry; Some consider Nisin as an antibiotic, but it would be necessary to carry out very rigorous clinical trials to consider it as such.

I agree with Dr. Karen

Hi Mushtak,

it's simple but very reliable for small studies as we known. there is two types of electrophoresis, agarose gel electrophoresis, and the second used by old AB sequencers, polyacrylamide gel. for what you asked, the gel (POPn) is included in capillaries by trough your samples will migrate. since your samples (bands or sequences) are fluorescent (4 dyes are used), they will be detected by the CCD camera at end of electrophoresis and interpretation of sizes will be done.

fred

Chronologically-

1) Alireza Mordadi - No quorum sensing is not a neither a signaling mechanism nor it regulates the deterministic factor for cells size determination or trigger for cell division. It is the mechanism which overall related with the population density in a particular environment. Basically it is radar homolog in bacteria. It might indirectly involve in the aforementioned, but not directly. Therefore, recommending quorum sensing (a wider topic) for a specific question is not supported.

2) Jay Sperry - since the discussion was not related to the biofilm but a bacterial growth in normal growth protocols, so I tried to relate the your previous comment to the context. And again it does not fit well, as you mentioned only one bacteria without mentioning any biofilm formation. "E. Faecalis forms biofilm after 14-15 days of incubation"(PMID: 27095620). So, what happens meanwhile, do cells divide or they remain in lag phase? And if we talk about E. faecalis biofilms, "SEM showed E. faecalis growth at all times" (PMID: 27095620).

Nevertheless, biofilms are special microbial structures, where the conventional growth curve does not applies. Therefore, involving biofilms in this discussion is like comparing apples and pears.

* I agree with your comment that bacteria might have different physiological growth phases, and it has been considered previously in the discussion above. Each species has its own growth rate and ofcourse cells requires time, a certain cell size and mass to initiate the division. But should we call it lag phase?

You can use Soybean casein digest agar for total aerobic microbial count. For total yeast and mould you can use Sabouraud dextrose agar.

To test E.coli in sample, first add pretreated -incubated sample to sterile macconkey broth and after further incubation subculture on Macconkey agar.

For salmonella, add pretreated-incubated sample to RVSE broth and after incubation subculture on XLD agar plate.

Standard test procedure described in USP chapter 61 and 62. Refer this.

Thanks a lot prof. Jawad K. Abood Al-Janabi

I am very thankful for your answer and your important suggestions,

I will put you microscopic photos for Byssochlamys spectabilis here.

Thanks again

Use gluteraldehyde or formaldehyde (1-5%) as cross linker. they undergo Schiff base reaction (CHO) of gluteraldehyde combine with NH2 group of protein). This reaction is only stable in alkaline condition, While at acidic condition use any stablizer to stable Schiff base reaction.

Nanodrop is great for routine PCR work but I would suggest that you use fluorometric quantification (Quantus) for sensitive applications such as high-throughput sequencing. The dye used in fluorometers selectively bind to dsDNA and it can give a more accurate quantification, especially in lower concentrations (if I remember correctly, <40ng/uL). Nanodrop measures all nucleotides so you can, in principle, read low-quality DNA such as sheared fragments from harsh extraction protocols. For regular PCR this is fine though.

In our lab, we use spectrophotometric methods for routine PCR as it is cheap. I have had experience with high Nanodrop readings but when I checked using agarose gels, the DNA were mostly sheared, likely due to over-beating of the starting material during lysis.

When it comes to high-throughput sequencing applications though, quantity AND quality/purity of DNA is of utmost importance so we opt for the Qubit or Quantus systems. It can be expensive (that's why we still use the Nanodrop) but I think it is worth it.

Bacteria can be engineered to produce electricity, according to many research. In the future, this kind of method could be used to help clean water in wastewater treatment plants.

It's a bit of an open question, could you be more specific? How much detail do you want? How much experience do you have? Have you ever identified compounds using GCMS? Who is guiding you in your lab?

I my opinion identifying compounds using GCMS is a trade, you learn it on the job by doing it many time and learning it from someone who knows what he/she is doing. You don't learn it from reading some publications, they can help but if nobody is guiding you in your laboratory, you will make mistakes. If you research is for a client or a publication then get help from someone in your lab who is experienced.

Identifying compounds is much more then running a NIST search and accepting the top compound if the match >80, still you see this kind of bad science in many publication.

Regards, Alex

Thanks Irene

Dear all,

thanks for all your comments and Khalid for your article! Finally, I found some time to take a closer look at the topic and this is what I can tell you so far:

Most monomeric RNAPs (like T7 RNAP) as well as DNAPs are members of a structural superfamily resembling an "half open" right hand sharing strong similarities in active site architecture and reaction mechanism. Substrate (i.e. r/dNTP) discrimination for DNAPs is accomplished by a "steric gate" (i.e. a bulky amino acid residue that would clash with the 2'OH-group of incoming rNTPs; e.g. E615 in Taq-pol., E480 in T7DNAP or E710 in Klenow Fragment) while RNAPs positively select for the 2'OH via hydrogen bonding (e.g. with Y639 in T7RNAP).

Accordingly, T7RNAP Y639F readily incorporates dNTPs into RNA transcripts but shows significantly decreased activity in vitro when only the 4 dNTPs are present (Sousa et al 1995). Likewise in 1995, Kostyuk et al reported that S641 would act as a "substrate specificity switch" and exchanging it with Ala would turn the T7 RNAP into an efficient DNAP. However, this was not reproduced by Huang and coworkers as stated in their 1997 paper and also the Moscow group around Kochetkov did not mention these earlier results about the S641A single mutant in their later works. So, I guess, unfortunately this would have been too convenient for us to be true and there was some sort of DNA-pol impurity in the sample or whatsoever. Anyhow, S641 apparently has no influence on rNTP/dNTP discrimination.

In the following years significant effort was made to generate polymerases that could produce nucleic acids containing non-canonical NTPs for uses in the clinic or as biosensors (e.g. as described in the paper Khalid provided).  These approaches, however, always aimed at engineering polymerases to tolerate bulkier substituents at the ribose's C2 position instead of tuning specificity towards dNTPs.

Perhaps one should try to use a directed evolution method to find appropriate enzyme variants or maybe it would be enough to combine the Y639F mutation with a deliberately introduced steric gate since T7RNAP only has a glycine at the respective position.

Please let me know your thoughts or if you know whether somebody ever tried to generate this double mutant.

If you are interested in this topic especially because you are fascinated by the notion of primerless DNA-synthesis then please read:

PS: You can find a list with all the relevant papers I could discover attached to this post.

@Madhukar Baburao Deshmukh and @Leena Rao: bacterial urea hydrolysis is one such reaction which produces alkaline pH. Are there any other similar reactions which will produce alkaline pH?

U can start with the evaluating the inhibition of enzyme activity to see the effect of the drug. Once you confirm inhibition, u should check the binding of drug to the enzyme using ITC, CD, fluorescence anisotropy/polarization etc. u can also perform docking studies of the drug on the structure/model of the protein followed by molecular dynamic simulation in order to understand the structural flexibility and dynamics of the protein upon drug binding. check some recent papers from our group, they may be helpful for your work.

Me as well,,,love me some over the counter Metatitanic acid (H2TiO3) ! Mess/me if you find some.

Occurrence of a complex of plant species in any geographic location is an indication of the adaptability of the species in that complex to the prevalent conditions in the location being studied. Adaptability, in turn, is an expression of genes for survival and reproduction  in the location. Initial lot of plants in this population might have come from diverse locations by way of dispersal or migration through various means available. Together, migration and survival make-up the adaptive radiation. At this stage, the change in genes in any species in the complex may not be much different from those prevailing in the parent populations from which they are derived. With advancing time and concurrent changes in climate, the genes for adaptation may change by way of recombining with related species in the vicinity, by mutation or by both. This results in a significant change in the expression of these genes in the next generation that may lead one to consider that new species are emerging. Sometimes, related species may interbreed and the resulting hybrids may tetraploidize as happened in the evolution of several crop species like coffee, wheat etc. In these cases speciation events are more clearly discernible.

You can used N2 gaz, with  specific conditions

you can use stainless steel bead with  sonication 

Hi Pawan,

Interesting topic. You may find these manuscripts and websites relevant:

Microbial biosensors

by Y Lei - ‎2006 - ‎

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Microbial Biosensors: Engineered Microorganisms as the Sensing ...

https://www.ncbi.nlm.nih.gov › NCBI › Literature › PubMed Central (PMC)

by M Park - ‎2013

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An impedimetric biosensor for real-time monitoring of bacterial growth ...

by YH Kim - ‎2009

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What file do you have?

If it's a fastq, count the number of lines and divide by four. If it's a sam/bam file use samtools to get the information.

If it were so simple, everybody would be doing it.

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Hi Emad. For in vitro mutagenicity tests it is best to follow the OECD guidelines attached below. I am working on different types of Fe nanoparticles and it is a good choice for in vitro mutagenicity studies. One drawback is the choice of concentrations which need cumbersome dosage screening experiments as Fe plays an active role in  cellular metabolism. So higher concentrations of Fe poses significant mutagenicity. Ag and Zn are known to be toxic and mutagenic in different biological systems. Our group has been actively working on them and they are the best nanoparticles to work on in vitro toxicity. Ag being an antibacterial nanoparticle induces genotoxicity and cell death and Zn at high concentrations causes DNA damage.

1. Yes you can use the same approach. MOI is simply the ratio of phage (pfu) to bacteria (cfu) e.g. an MOI of 1 is an equal concentration of both and an MOI of 10 is a 10-fold excess of phage. Dilutions of the phage can be made in any standard phage buffer usually containing divalent cations e.g. Ca2+ or Mg2+.

2. You can use whatever volumes you want to use for this so long as the ratio is correct. Ideally you would start with a concentrated phage stock to avoid excessive dilution of the bacteria, which will affect your yield. If your aim is just to propagate the phage then total volume is not important, however, is you are testing infection frequencies at different MOIs then you should ensure the conditions are identical between experiments e.g. 100 ul bacteria (OD600=0.5) plus 100 ul phage diluted to desired concentration.

3. Infections can be incubated either shaking or stationary

Dear Tijjani,

The following publications cover the answer to your question. The first publication is a review article describes the pretreatment process of lignocellulosic wastes to improve ethanol and biogas production. The second publication is a paper that describes the pretreatment process of orange peel by leaching of limonene for improving biogas production.

Mohammad J. Taherzadeh1,* and Keikhosro Karimi1,2

Author information ► Article notes ► Copyright and License information ►

This article has been cited by other articles in PMC.

 

Go to:

Abstract

Lignocelluloses are often a major or sometimes the sole components of different waste streams from various industries, forestry, agriculture and municipalities. Hydrolysis of these materials is the first step for either digestion to biogas (methane) or fermentation to ethanol. However, enzymatic hydrolysis of lignocelluloses with no pretreatment is usually not so effective because of high stability of the materials to enzymatic or bacterial attacks. The present work is dedicated to reviewing the methods that have been studied for pretreatment of lignocellulosic wastes for conversion to ethanol or biogas. Effective parameters in pretreatment of lignocelluloses, such as crystallinity, accessible surface area, and protection by lignin and hemicellulose are described first. Then, several pretreatment methods are discussed and their effects on improvement in ethanol and/or biogas production are described. They include milling, irradiation, microwave, steam explosion, ammonia fiber explosion (AFEX), supercritical CO2 and its explosion, alkaline hydrolysis, liquid hot-water pretreatment, organosolv processes, wet oxidation, ozonolysis, dilute-and concentrated-acid hydrolyses, and biological pretreatments.

For more details, see attached file.

Rachma Wikandari,1 Huong Nguyen,2 Ria Millati,3

Claes Niklasson,2 and Mohammad J. Taherzadeh1

Limonene is present in orange peel wastes and is known as an antimicrobial agent, which impedes biogas production when digesting the peels. In this work, pretreatment of the peels to remove limonene under mild condition was proposed by leaching of limonene using hexane as solvent. The pretreatments were carried out with homogenized or chopped orange peel at 20–40∘ C with orange

peel waste and hexane ratio (w/v) ranging from 1 : 2 to 1 : 12 for 10 to 300 min. The pretreated peels were then digested in batch reactors for 33 days. The highest biogas production was achieved by treating chopped orange peel waste and hexane ratio of 12 : 1 at 20∘ C for 10 min corresponding to more than threefold increase of biogas production from 0.061 to 0.217 m3 methane/kg VS. The

solvent recovery was 90% using vacuum filtration and needs further separation using evaporation. The hexane residue in the peel had a negative impact on biogas production as shown by 28.6% reduction of methane and lower methane production of pretreated orange peel waste in semicontinuous digestion system compared to that of untreated peel.

For your question you may need to read the first publication (review), however, I attached the second publication that might be helpful for you as well.

Hoping this will be helpful,

Rafik

compare pure pullulan and your sample, assuming that pure pullulan is totaly converted glucose

Dear BiveK,

I would definitely recommend MS. By MS you can identify specific proteins that are highly expressed in you cancer cell lines, or even directly compare the MS spectrum between the cancer cell lines and normal cell lines. After identifying these possible peptides you would proceed with tandem MS to determine the specific sequence of each of the previously identified peptides. 

Here is some relevant literature in the matter:

Protein Sequencing and Identification Using Tandem Mass Spectrometry by Michael Kinter and Nicholas E. Sherman, Wiley

Teen and Mann (2004) The ABC’s (and XYZ’s) of peptide sequencing. Nature Reviews 5:699-711.

Kellie et al. (2010) The emerging process of top down mass spectrometry for protein analysis: biomarkers, protein-therapeutics, and achieving high throughput. Mol Biosyst. 6:1532-1539.

Hope this helps you,

Best,

Rita Pedrosa

If you want solve this problem, you need to test if that is the case. What you need to do is to do expression trials at different temperatures. Usually you can get more full-length at a lower temperature.

Most protein degradation occur at the C-terminus, if you fuse a affinity tag at the C-terminus of your protein, you might be able to purify the full length version even with some degradation. However, this kind trouble-shooting  is always in a style of "try and error",  you can only work it out after trying with different approaches.

I have identified some industrial proteins from Bacillus licheniformis and Bacillus subtilis (both gram positive) which include alpha-amylase, cellulase, Bacitracin synthetase, signal peptidase, leucine aminopeptidase precursor proteins and a host of others. 

I could be associated with differential pressure. There may be a malfunction of the steam trap during the filter sterilization process in the fermenter and this causes a pressure drop that is capable of piercing the mambrana. This type of membrane may fail under mechanical stress.

The best process to do that  is nanofiltration

Hi Bharat Bhushan Negi

I've read on Sadasivam, S. & Manickam, A. 1996. Biochemical Methods. New Age International. Page 13 - 14.

It was an easy method of activity determination and published in 1987 by Ghose, using cellulose filter paper, high purity. For simplicity is adopted it as a parameter. The cellulose in the filter requires endoglucanases, exoglucanases and beta-glucosidases (three types of cellulase), thus, measuring the total cellulase activity.

This article is:

Plants do uptake ions as dissolved in water vias specific transporters. They may increase the concentration by sequestering into organelles such as vacuoles. They do also form complexes with others once they are inside the cell environment. Concept of Micro Nano Particles are not yet been adapted in plant related literatures (As far as i know). For time being you can assume MNP as ionic form of metal in water. 

Co culture could be developed both by different species or the same species. The main difference is that mixed cultures are usually present in some environments such as activated sludge. they are extracted and bring to the labs, but co cultures are developed synthetically using different microorganisms which are grown in the same medium.

Dear  Arushdeep

From NCBI  GenBank view,  click "Highlight features" in the extreme right of the sequence panel in question.  You may as well click "gene" and go to the graphical view where you will be able to see the regions from "Tools."

However,  the best thing would be is to use prediction servers such as the one hosted in DTU: http://www.cbs.dtu.dk/services/Promoter/

Best,

Prash 

dear mohamed, i am glad of being of some help to you

Please find the answer for your question from the link below:

Dear Carlos,

It depends on what kind of modulation of the ETC you want to see? If you want to observe dynamic changes in mitochondria membrane potential, which is of course tightly linked to ETC function, then yes it is a good method. For this a final concentration of 10ug/mL should work well as a dequench mode concentration(30 min incubation at RT). You could use verapamil in order to block MDR transporters but I don`t think uptake into mitochondria will be severely affected at this concentration. Are you limited to a plate reader assay or could you use microscopy or polarographic oxygen consumption measurements? Hope it helps! 

Unfortunately, you have excluded two of the best suited metals for making electrodes.

Aluminum is a cheap metal, which is readily available in convenient sheets of many sizes. But Al oxidizes fairly quickly, which may cause changes in the ability of its surface to conduct current repeatedly as it is used. You may have to removed oxidized layer before using to avoid this problem.

You could also consider making your electrodes with a metal of high electrical conductivity and then getting it gold plated, which should not be nearly as expensive as using sheets of gold.

Hello, Greetings. Kindly follow the attched files for your reference.

Hi Kathir,

In the Glucose/Lactose system, the synthesis of T7 RNA polymerase will be repressed as there is glucose available as the primary source for metabolism. Only when the glucose is exhausted the lactose can drive the T7 RNA polymerase synthesis,  by promoting the transcription of the target mRNA of the gene cloned in the plasmid vector. 

Coming to the point, even if you express the target protein with lactose at lower temperature, some proteins forms inclusion bodies. It is based on the hydrophobic nature of the protein.

But definitely the amount of enzyme activity will not be increased by expressing at lower temperature. Because the rate of protein expression is lesser at lower temperature, so the amount of protein expressed also will be lesser than expressing at higher temperature.

Hi Lisle, what is the protein composition of the CF-ECM you are looking for?

If you try searching a database like KEGG for your gene of interest, the taxonomy will list all of the genomes in which the merA gene have been identified. I believe that a merA gene has been annotated in the genome of L. crispatus.

Hi Mohsen,

I have seen some papers published using 35S promoter to drive a gene in some fungi. If you cannot find documentation about transgenesis on basidimycetes (your experiment material), you can first try some transient assay using 35S promoter and a GFP gene (35S::gfp) to see if the 35S promoter can drive the gfp gene and glow in your fungus tissue. You can try a quick bombardment experiment. You can also use other screenable genes, such as gus gene.

Please refer this link:

Yes, organic solvents will influence the tertiary (and secondary) structure of proteins. I have once studied the effect of hexafluorisopropanol (HFIP), which is miscible with water in low and high concentrations (but inbetween is a miscibility gap).

HFIP reportedly enhances the alpha helicity of the secondary structure, and the

tertiary structure of proteins is disordered by this solvent.

I have not used toluene and I do not know what it does to the alpha helicity of the secondary structure, but I would assume it to disorder the tertiary structure as well. As long as it does not precipitate, that should not be a problem, unless you want to do biological assays with it afterwards.

Hope this answers your question

You need to look for the root cause for high bioburden. Is it coming form environment / raw material? Are cleanroom procedures stringently followed? If E.coli is the problem, is it coming from your water? You need to confirm quality of your water purification and sterilization system also. Post-production reduction will require harsh treatments; if it is a serious problem, you may try low level (~5kGy) gamma irradiation if the product can sustain.

actually It's depend on the type of your unknown ? is it solid , liquid or gas phase . organic , inorganic or bio-organic molecule .clarify the physical properties of the unknown that will help. generally .FT-IR + H-NMR+ C-NMR , HPLC, GC and Mass analysis ...etc all of these technique will help to elucidation your unknown product .

hello!

I agree with former answers - stalked ciliates are normally found in starting phases of ASWWTPs, in concentrations ranging from 10E4 to 10E5 / liter.

But they are also well adapted under transient conditions, i.e. sludge losses, hydraulic transient overload or intermittent low recirculating flow. For instance, they may be encountered in concentrations over 10E6 / liter as soon as one day after a recirculating pump breakdown.

Furthermore, stalked ciliates genera differ strongly along with other parameters like effluent origin or aeration pattern, so it would be a valuable information to identify the genera (refer to the old Curds identification key, more recently traduced in italian by Madoni, or to the excellent "how to know the protozoa" by Jahn and Bovee).

For more details refer to following papers and especially the last one (I strongly apologize for these papers are very old and written in french...):

DRAKIDÈS C., 1977. Étude d'une biocénose en conditions limitantes. Cas de l'adaptation d'une boue activée à un effluent industriel (monopropylène- glycol en présence de Ca Cl2). La Technique de l'Eau et de l'Assainissement, 365, 21-32.

DRAKIDÈS C., 1978. L'observation microscopique des boues activées appliquée à la surveillance des installations d'épuration: technique d'étude et interprétation. Techniques et Sciences Municipales, 2 - 78, 85-98.

DRAKIDÈS C., 1980. La microfaune des boues activées. Étude d'une méthode d'observation et application au suivi d'un pilote en phase de démarrage. Water Research, 14, 1199-1207.

I love this question!

If you love Plants + Biotechnology, I highly recommend you to take both courses. As a Biotechnology Engineering student, I had my first course called "Botanicals and plants physiology" which was the first step to understand the plants and their metabolism, which was a very important base to do some more understanding before applying that knowledge with the biotechnology area. After that, I had a course called "Plants Biotechnology" which as the other comments says, you can deeply understand the callus formation, the strategy and protocols to do in vitro propagation, and how to apply all your knowledge to use plants as tools to solve problems.

I hope you enjoy all courses you do, they're all useful :) !