Jing Yuan

Zhengzhou University, Zhengzhou, Henan Sheng, China

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Publications (26)91.12 Total impact

  • Article: Rapid detection of Trichinellaspiralislarvae in muscles by loop-mediated isothermal amplification.
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    ABSTRACT: Trichinellaspiralisis a tissue-dwelling nematode parasite. A loop-mediated isothermal amplification (LAMP) assay was developed and validated for the sensitive and rapid detection of T. spiralis larvae in muscle samples. Sixteen sets of primers were designed to recognize distinct sequences of a conserved gene, a 1.6kb repetitive element of the Trichinella genome. One set of primers was selected as the most appropriate for rapid detection. The specificity and sensitivity of the primers in LAMP reactions for T. spiralis larvae and muscle samples of mice infected with T. spiraliswere determined. Another 10 heterologous parasites were selected for specificity assays. The results showed that target DNA was amplified and visualized by monitoring turbidity and adding calcein detection methods within 70 min at an isothermal temperature of 63°C. The sensitivity of LAMP with the detection limit of 362 fg/μl was >10 times higher than that for PCR. The designed primers had a good specificity. No cross-reactivity was found with the DNA of any other parasites. The assay was able to detect T. spiralis in all mouse muscle samples infected with 10 T. spiralis larvae on day 20 p.i. We believe this is the first report regarding the application of the LAMP assay for detection of T. spiralis larvae in muscle samples from experimentally infected mice. This method demonstrates a potentially valuable means for the direct detection of T. spiralis larvae in meat inspection.
    International journal for parasitology 11/2012; · 3.39 Impact Factor
  • Article: Shigella flexneri T3SS effector IpaH4.5 modulates the host inflammatory response via interaction with NF-κB p65 protein.
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    ABSTRACT: Shigella species possess a type III secretion system (T3SS), which is required for human infection and that delivers effector proteins into target host cells. Here, we show that the effector, IpaH4.5 dampens the pro-inflammatory cytokine response. In both the Sereny test and a murine lung infection model, the Shigella ΔipaH4.5 mutant strain caused more severe inflammatory responses and significantly induced higher pro-inflammatory cytokine levels (MIP-2 and TNF-α) in the lung homogenates of wild type-infected mice. Moreover, there was a 3-fold decrease in bacterial colonization of the mutant compared to the WT and ΔipaH4.5/ipaH4.5-rescued strains. Yeast two-hybrid screening showed that IpaH4.5 specifically interacts with the p65 subunit of NF-κB. Ten truncated versions of IpaH4.5 and p65 spanning different regions were constructed and expressed to further map the IpaH binding sites with p65. The results revealed that the p65 region spanning amino acids 1-190 of p65 interacted with the IpaH4.5/1-293 N-terminal region. In vitro, IpaH4.5 displayed ubiquitin ligase activity toward ubiquitin and p65. Furthermore, the transcriptional activity of NF-κB was shown to be inhibited by IpaH4.5 utilizing a dual-luciferase reporter gene detection system containing NF-κB promoter response elements. Thus, we conclude that the IpaH4.5 protein is an E3 ubiquitin ligase capable of directly regulating the host inflammatory response by inhibiting the NF-κB signaling pathway.
    Cellular Microbiology 10/2012; · 5.46 Impact Factor
  • Article: High Rate of New Delhi Metallo-β-Lactamase 1-Producing Bacterial Infection in China.
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    ABSTRACT: New Delhi Metallo-β-Lactamase 1 (NDM-1) is spreading worldwide and represents a major threat to health. To investigate the infection status and the species distribution of NDM-1-producing bacteria in China, a total of 10273 clinical fresh faecal samples of separate individuals were collected and analysed. Our results showed that the total infection rates of NDM-1-producing intestinal bacteria in clinical patients were 14.8%.
    Clinical Infectious Diseases 09/2012; · 9.15 Impact Factor
  • Article: Parallel gene loss and acquisition among strains of different Brucella species and biovars.
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    ABSTRACT: The genus Brucella is divided into six species; of these, B. melitensis and B. abortus are pathogenic to humans, and B. ovis and B. neotomae are nonpathogenic to humans. The definition of gene loss and acquisition is essential for understanding Brucella's ecology, evolutionary history, and host relationships. A DNA microarray containing unique genes of B. melitensis Type strain 16MT and B. abortus 9-941 was constructed and used to determine the gene contents of the representative strains of Brucella. Phylogenetic relationships were inferred from sequences of housekeeping genes. Gene loss and acquisition of different Brucella species were inferred. A total of 214 genes were found to be differentially distributed, and 173 of them were clustered into 15 genomic islands (GIs). Evidence of horizontal gene transfer was observed for 10 GIs. Phylogenetic analysis indicated that the 19 strains formed five clades, and some of the GIs had been lost or acquired independently among the different lineages. The derivation of Brucella lineages is concomitant with the parallel loss or acquisition of GIs, indicating a complex interaction between various Brucella species and hosts.
    The Journal of Microbiology 08/2012; 50(4):567-74. · 1.10 Impact Factor
  • Article: Proteomic analysis of clinical isolate of Stenotrophomonas maltophilia with blaNDM-1, blaL1 and blaL2 β-lactamase genes under imipenem treatment.
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    ABSTRACT: The co-occurrence of L1 and AmpR-L2 with bla(NDM-1) gene with an upstream 250-bp promoter was detected in a clinical isolate of Stenotrophomonas maltophilia DCPS-01, which was resistant to all β-lactams and sensitive only to colistin and fluoroquinolones. To investigate expression of resistance genes and the molecular mechanisms of bacteria resistance to carbapenems, proteomic profiles of the isolate was passaged with and without the drug by using 2D-PAGE. The results showed that 33 genes exhibiting a ≥3-fold change were identified as candidates that may help S. maltophilia survive drug selection. Strikingly, L1 was expressed more highly in cells grown with imipenem, and the abundant NDM-1 further increased, while very little L2 was detected even following induction. Specific activities for β-lactamase revealed that L2 remained at constitutive low levels (10.6 U/mg), while L1 and NDM-1 showed clear activity (69.8 U/mg). Our data support that imipenem could specifically and reversibly induce L1 and NDM-1, which together played key roles in drug resistance in DCPS-01. Although NDM-1 mediated resistance to carbapenems has been found in very few cases, to our knowledge, this is the first proteomics research of S. maltophilia with NDM-1, giving very broad-spectrum antibiotic resistance profiles.
    Journal of Proteome Research 06/2012; 11(8):4024-33. · 5.11 Impact Factor
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    Article: Improved adhesive properties of recombinant bifidobacteria expressing the Bifidobacterium bifidum-specific lipoprotein BopA.
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    ABSTRACT: Bifidobacteria belong to one of the predominant bacterial groups in the intestinal microbiota of infants and adults. Several beneficial effects on the health status of their human hosts have been demonstrated making bifidobacteria interesting candidates for probiotic applications. Adhesion of probiotics to the intestinal epithelium is discussed as a prerequisite for colonisation of and persistence in the gastrointestinal tract. In the present study, 15 different strains of bifidobacteria were tested for adhesion. B. bifidum was identified as the species showing highest adhesion to all tested intestinal epithelial cell (IEC) lines. Adhesion of B. bifidum S17 to IECs was strongly reduced after treatment of bacteria with pronase. These results strongly indicate that a proteinaceous cell surface component mediates adhesion of B. bifidum S17 to IECs. In silico analysis of the currently accessible Bifidobacterium genomes identified bopA encoding a lipoprotein as a B. bifidum-specific gene previously shown to function as an adhesin of B. bifidum MIMBb75. The in silico results were confirmed by Southern Blot analysis. Furthermore, Northern Blot analysis demonstrated that bopA is expressed in all B. bifidum strains tested under conditions used to cultivate bacteria for adhesion assays. The BopA gene was successfully expressed in E. coli and purified by Ni-NTA affinity chromatography as a C-terminal His6-fusion. Purified BopA had an inhibitory effect on adhesion of B. bifidum S17 to IECs. Moreover, bopA was successfully expressed in B. bifidum S17 and B. longum/infantis E18. Strains overexpressing bopA showed enhanced adhesion to IECs, clearly demonstrating a role of BopA in adhesion of B. bifidum strains. BopA was identified as a B. bifidum-specific protein involved in adhesion to IECs. Bifidobacterium strains expressing bopA show enhanced adhesion. Our results represent the first report on recombinant bifidobacteria with improved adhesive properties.
    Microbial Cell Factories 06/2012; 11:80. · 3.55 Impact Factor
  • Article: Accessing the inaccessible: molecular tools for bifidobacteria.
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    ABSTRACT: Bifidobacteria are an important group of the human intestinal microbiota that have been shown to exert a number of beneficial probiotic effects on the health status of their host. Due to these effects, bifidobacteria have attracted strong interest in health care and food industries for probiotic applications and several species are listed as so-called "generally recognized as safe" (GRAS) microorganisms. Moreover, recent studies have pointed out their potential as an alternative or supplementary strategy in tumor therapy or as live vaccines. In order to study the mechanisms by which these organisms exert their beneficial effects and to generate recombinant strains that can be used as drug delivery vectors or live vaccines, appropriate molecular tools are indispensable. This review provides an overview of the currently available methods and tools to generate recombinant strains of bifidobacteria. The currently used protocols for transformation of bifidobacteria, as well as replicons, selection markers, and determinants of expression, will be summarized. We will further discuss promoters, terminators, and localization signals that have been used for successful generation of expression vectors.
    Applied and environmental microbiology 05/2012; 78(15):5035-42. · 3.69 Impact Factor
  • Article: Fructose uptake in Bifidobacterium longum NCC2705 is mediated by an ATP-binding cassette transporter.
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    ABSTRACT: Recently, a putative ATP-binding cassette (ABC) transport system was identified in Bifidobacterium longum NCC2705 that is highly up-regulated during growth on fructose as the sole carbon source. Cloning and expression of the corresponding ORFs (bl0033-0036) result in efficient fructose uptake by bacteria. Sequence analysis reveals high similarity to typical ABC transport systems and suggests that these genes are organized as an operon. Expression of FruE is induced by fructose, ribose, or xylose and is able to bind these sugars with fructose as the preferred substrate. Our data suggest that BL0033-0036 constitute a high affinity fructose-specific ABC transporter of B. longum NCC2705. We thus suggest to rename the coding genes to fruEKFG and the corresponding proteins to FruE (sugar-binding protein), FruK (ATPase subunit), FruF, and FruG (membrane permeases). Furthermore, protein-protein interactions between the components of the transporter complex were determined by GST pulldown and Western blot analysis. This revealed interactions between the membrane subunits FruF and FruG with FruE, which in vivo is located on the external side of the membrane, and with the cytoplasmatic ATPase FruK. This is in line with the proposed model for bacterial ABC sugar transporters.
    Journal of Biological Chemistry 11/2011; 287(1):357-67. · 4.77 Impact Factor
  • Article: Proteomics analysis of Bifidobacterium longum NCC2705 growing on glucose, fructose, mannose, xylose, ribose, and galactose
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    ABSTRACT: To investigate the molecular mechanisms underlying carbohydrate uptake and connected metabolic pathways of Bifidobacterium longum NCC2705, the proteomic profiles of bacteria grown on different carbon sources including glucose, fructose, mannose, xylose, ribose, and galactose were analyzed. Our results show that all sugars tested were catabolized via the bifid shunt. Sixty-eight proteins that exhibited changes in abundance of threefold or greater were identified by MS. A striking observation was the differential expression of proteins related to the pyruvate metabolism. Further analysis of acetic acid and lactic acid in the culture supernatants by HPLC at the end of fermentation showed that more lactic acid was produced during growth on fructose, ribose, xylose, galactose and more acetic acid was produced during the fermentation of glucose and mannose. Growth experiments revealed that B. longum NCC2705 preferentially used fructose, ribose, xylose, and galactose with higher growth rates over glucose and mannose. Furthermore, five proteins (GroEL, Eno, Tal, Pgm, and BL0033) exhibited clear phosphorylation modifications at serine and/or tyrosine residues. BL0033, a component of an ATP-binding cassette (ABC) transporter, was significantly more abundant in bacteria grown on fructose and, to a lesser extent, ribose and xylose. RT-PCR analysis revealed that all genes of the ABC transporter are induced in the presence of these sugars suggesting that BL0033, BL0034, BL0035, and BL0036 constitute an ABC transporter with fructose as preferred substrate.
    Proteomics 05/2011; 11(13):2628 - 2638. · 4.43 Impact Factor
  • Article: Development of an extended multilocus sequence typing for genotyping of Brucella isolates.
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    ABSTRACT: By amplifying and sequencing longer sequences, an extended multi locus sequence typing (EMLST) theme was developed for Brucella. 61 isolates were genotyped by the EMLST with increased resolution. This strategy could be extended to other bacteria to improve MLST genotyping resolution without additional loci.
    Journal of microbiological methods 05/2011; 86(2):252-4. · 2.43 Impact Factor
  • Article: The 16MΔvjbR as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection.
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    ABSTRACT: Brucellosis brings great economic burdens for developing countries. Live attenuated vaccines are the most efficient means for prevention and control of animal Brucellosis. However, the difficulties of differentiating of infection from vaccine immunization, which is essential for eradication programs, limit their applications. Therefore, the development of a vaccine that could differentiate infection from immunization will overcome the limitations and get extensive application. VjbR is a quorum sensing regulator involving in Brucella's intracellular survival. The vjbR∷Tn5 mutants have been proven effective against wild type strain challenge, implying its possibility of use in vaccine candidate development. To further evaluate this candidate gene, in the present study, the antigenicity of purified recombinant VjbR protein was analyzed. Antibodies to Brucella melitensis VjbR could be detected in sera from patients and animals with brucellosis but not in control ones, implying the potential use of this protein as a diagnostic antigen. Then a vjbR mutant of B. melitensis 16M was constructed by replacing the vjbR with kanamycin gene. The mutant showed reduced survival in macrophage and mice. Vaccination of BALB/c mice with 16MΔvjbR conferred significant protective immunity against B. melitensis strain 16M challenges, being equivalent to which induced by the license vaccine Rev.1. The vjbR deletion mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon and interleukin-10. The most importance is that, the use of vjbR mutants as vaccines in association with diagnostic tests based on the VjbR antigen would allow the serological differentiation between infected and vaccinated animals. These results suggest that 16MΔvjbR is an ideal live attenuated vaccine candidate against B. melitensis and deserves further evaluation for vaccine development.
    Veterinary Microbiology 04/2011; 151(3-4):354-62. · 3.33 Impact Factor
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    Article: [Applications and perspectives of DNA stable-isotope probing in metagenomics: a review].
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    ABSTRACT: DNA stable-isotope probing (DNA-SIP) is a recently developed method with which the incorporation of stable isotope from a labeled substrate is used to identify the function of microorganisms in the environment. The technique has now been used in conjunction with metagenomics to establish links between microbial identity and particular metabolic functions. The combination of DNA-SIP and metagenomics not only permits the detection of rare low-abundance species from metagenomic libraries but also facilitates the detection of novel enzymes and bioactive compounds. We summarize recent progress in SIP-metagenomic techniques and applications and discuss prospects for this combined approach in environmental microbiology and biotechnology.
    Sheng wu gong cheng xue bao = Chinese journal of biotechnology 04/2011; 27(4):539-45.
  • Article: Coexistence of blaNDM-1 with the prevalent blaOXA23 and blaIMP in pan-drug resistant Acinetobacter baumannii isolates in China.
    Clinical Infectious Diseases 03/2011; 52(5):692-3. · 9.15 Impact Factor
  • Article: Proteomic analysis of the Enterococcus faecalis V583 strain and clinical isolate V309 under vancomycin treatment.
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    ABSTRACT: To understand the molecular mechanisms of bacteria resistance to glycopeptides, we obtained proteomic profiles of vancomycin-resistant Enterococcus faecalis V583 (reference strain) and V309 (clinical isolate) passaged with and without the drug. The specificity and reversibility of vancomycin resistance genes induced in V583 and V309 were further studied over time. By semiquantitative RT-PCR of vancomycin-treated versus untreated samples of both strains, 28 (V583) or 20 (V309) up-regulated proteins, 8 (V583) or 6 (V309) down-regulated proteins, and 1 (V583) or 4 (V309) proteins with mobility changes in 2-DE gel analysis were identified. Some of these proteins have known vancomycin resistance functions or are related to virulent factors, stress, metabolism, translation, and conjunction, which would help Enterococcus survive under drug selection. Vancomycin induced specifically and reversibly VanA, VanX, VanB, and VanXB. Notably, 6 proteins (Pgm, Ldh, Gap-2, RpsB, EF2076, and sex pheromone cAD1 precursor lipoprotein) exhibited clear post-translational modifications. Vancomycin induced phosphorylation of Ser/Thr in Ldh, Gap-2, and sex pheromone cAD1 precursor lipoprotein (EF3256), newly identified here as enterococcal phosphoproteins. Our data suggest that phosphorylated EF3256 is normally active in E. faecelis, whereas EF3256-P together with oppA-like protein may play a key role in the regulation of pheromone and transmission of conjugation plasmids.
    Journal of Proteome Research 04/2010; 9(4):1772-85. · 5.11 Impact Factor
  • Article: The type IV secretion system affects the expression of Omp25/Omp31 and the outer membrane properties of Brucella melitensis.
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    ABSTRACT: The type IV secretion system (T4SS) contributes to Brucella intracellular survival through its effector proteins. Comparative proteomic analysis showed that intracellular survival proteins are expressed differentially in a virB mutant. Interestingly, several outer membrane proteins (OMPs) are also differentially expressed, implying that T4SS might affect the OM properties of Brucella. To further evaluate the impact of T4SS on OM, in the present study, the OM proteomes were isolated and compared. Many more products of OMPs, particularly different products of the Omp25/Omp31 family, were found to be altered in the virB mutant. The transcription profiles of Omp25/Omp31 were different from those of their protein products, implying their regulation by virB at both transcriptional and post-transcriptional levels. The virB mutant aggregates at a high cell density and produces exopolysaccharide, a phenotype resembling that of the vjbR mutant. The virB mutant showed increased sensitivity to polymyxin B and decreased survival under oxidative, high-salt and high-osmolarity stresses, indicating drastic membrane alterations. These results indicated that in addition to being an effector protein secretion system, T4SS affects OM properties that might be important for the adaptation of Brucella to both in vitro and in vivo hostile environments.
    FEMS Microbiology Letters 02/2010; 303(1):92-100. · 2.04 Impact Factor
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    Article: Comparative proteomics analyses reveal the virB of B. melitensis affects expression of intracellular survival related proteins.
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    ABSTRACT: Brucella melitensis is a facultative, intracellular, pathogenic bacterium that replicates within macrophages. The type IV secretion system encoded by the virB operon (virB) is involved in Brucella intracellular survival. However, the underlying molecular mechanisms, especially the target proteins affected by the virB, remain largely unclear. In order to define the proteins affected by virB, the proteomes of wild-type and the virB mutant were compared under in vitro conditions where virB was highly activated. The differentially expressed proteins were identified by MALDI-TOF-MS. Forty-four down-regulated and eighteen up-regulated proteins which exhibited a 2-fold or greater change were identified. These proteins included those involved in amino acid transport and metabolism, lipid metabolism, energy production, cell membrane biogenesis, translation, post-translational modifications and protein turnover, as well as unknown proteins. Interestingly, several important virulence related proteins involved in intracellular survival, including VjbR, DnaK, HtrA, Omp25, and GntR, were down-regulated in the virB mutant. Transcription analysis of virB and vjbR at different growth phase showed that virB positively affect transcription of vjbR in a growth phase dependent manner. Quantitative RT-PCR showed that transcription of these genes was also affected by virB during macrophage cell infection, consistent with the observed decreased survival of the virB mutant in macrophage. These data indicated that the virB operon may control the intracellular survival of Brucella by affecting the expression of relevant proteins.
    PLoS ONE 02/2009; 4(4):e5368. · 4.09 Impact Factor
  • Article: [Proteomic analysis of Bifidobacteria longum strain NCC2705 grown on lactose and glucose].
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    ABSTRACT: Based on a proteomic reference map of the important probiotic organism Bifidobacteria longum NCC2705 constructed by our previous research, we compared the proteomic profiles of Bifidobacteria longum strain NCC2705 grown on lactose or glucose to identify the catabolic route allowing lactose fermentation. We considered the proteins differentially expressed if their relative volume deviated more than 3-fold with ImageMaster 2D Elite version 5.0 software. Interesting spots were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis, and phosphorylation analysis of proteins with mobility changes by Pro-Q Diamond Stain. The identified spots represent 31 protein entries, 14 up-regulated proteins, 17 down-regulated proteins. These identified proteins, which were hydrophilic proteins and their genes with CAI value above 0.5 represented the most abundant proteins, included key stress proteins, metabolism-related proteins, and proteins related to translation. Two proteins including Tal (BL0715, transaldolase, L3) and Pyk (BL0988, pyruvate kinase, G9) exhibited clear post-translational modification. Proteomic comparison of glucose- and lactose-grown cells revealed that lactose and glucose were catabolized via the same degradation pathway, and the rate of glucose assimilation was higher than that of lactose. Spot and protein analysis revealed that post-translational modifications might be common in these proteins. Pro-Q Diamond staining analysis revealed that lactose trigger Tal phosphorylation at 43 T /47 S, and inhibited Pyk phosphorylation at 65 S. These proteins were identified for the first time as bifidobacterial phosphoproteins.
    ACTA MICROBIOLOGICA SINICA 12/2008; 48(11):1451-8.
  • Article: [Comparative proteome analysis of Bifidobacterium longum NCC2705 grown on fructose and glucose].
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    ABSTRACT: To demonstrate the fructose metabolism pathway in Bifidobacterium Longum NCC2705 and to construct its fermentation model, we explored the comparative proteome cultivating the strain on glucose or fructose, based on a proteomic reference map of B. longum NCC2705 constructed earlier. Then, we used matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and electro-spray ionization tandem mass spectrometry (ESI-MS/MS) for differently expressed proteins identification. Furthermore, with semi-quantitative RT-PCR we determined the distinctively expressed proteins at the level of transcription. Proteomic comparison of glucose- and fructose-grown cells demonstrated much similarity. On the page of fructose there were all the enzymes and proteins that exist during the process of glucose degradation. We observed a greater variation of more than three-fold for the identified 9 spots representing 5 protein entries by MALDI-TOF MS. The sugar-binding protein specific to fructose (BL0033) and an ABC transporter ATP binding protein (BL0034) showed higher expression level from cells grown on fructose. It was also determined by semi-quantitative RT-PCR subsequently. BL0033 time course and concentration experiments showed that the induction time correlated to higher fructose concentration, and increased expression of BL0033. Fructose was catabolized via the same degradation pathway as glucose at the level of proteomics. BL0033 was induced by fructose. All results suggest that the uptake of fructose into the cell may be conducted by a specific ABC transport system, in which BL0033 and BL0034 as components might have played an important role.
    Sheng wu gong cheng xue bao = Chinese journal of biotechnology 09/2008; 24(8):1401-6.
  • Article: [HtpG protein of Shigella flexneri 2a strain 2457T evokes inflammatory response in mice].
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    ABSTRACT: To analyze the function of htpG of S. flexneri 2a 2457T, we constructed an htpG deletion mutant and a recovery mutant. gamma-Red recombination system was used to construct an htpG deletion mutant of S. flexneri 2a 2457T. In addition, a recover mutant was obtained by introducing a low-copy plasmid containing one copy of htpG gene into the deletion mutant. Then, the growth curves of wild-type strain, deletion mutant and recover mutant were measured. Some of biochemical tests were also investigated. Furthermore, the Sereny tests were performed to evaluate the virulence of these strains. No significant difference were observed among three strains. However, the titers of some inflammatory factors evoked by wild-type strain, deletion mutant and recovery mutant in intraperitoneal injected mice were quite different. These results suggest that the HtpG protein of Shigella flexneri 2a strain 2457T might be involved in the immunopathogenesis.
    ACTA MICROBIOLOGICA SINICA 08/2008; 48(7):905-10.
  • Article: [Recombinational cloning of large DNA fragments of bacterial chromosome by plasmid rescue].
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    ABSTRACT: To develop a new method to cline large DNA fragment by combining whole genome sequence information and the principle of plasmid rescue. First, we amplified a fragment downstream the region to be cloned and cloned the fragment into a suicide plasmid to construct targeting plasmid, which was then targeted into chromosome by homologous recombination. Then, genomic DNA was isolated, digested with appropriate enzyme, re-ligated and transformed into host bacteria to rescue the plasmid and the large DNA fragment. The rescued DNA fragment was sub-cloned into new plasmids for special purposes. Using this method, we successfully cloned the 11kb virB operon of Brucella and constructed complementary plasmid. The transcription of the disrupted virB operon was restored with the complementary plasmid, validating the feasibility of the strategy. This recombinational cloning strategy provides us a new method to modify large DNA fragment of bacteria.
    ACTA MICROBIOLOGICA SINICA 05/2008; 48(4):532-8.

Institutions

  • 2012
    • Zhengzhou University
      Zhengzhou, Henan Sheng, China
  • 2008–2012
    • Academy of Military Medical Sciences
      Tianjin, Tianjin Shi, China
    • Northwest A & F University
      • College of Food Science and Engineering
      Yangling, Shaanxi Sheng, China
  • 2006
    • Fuerkang Beijing Institute of Biotechnology
      Beijing, Beijing Shi, China
  • 2005
    • State Key Laboratory of Medical Genetics of China
      Changsha, Hunan, China