Demin Jin

Hebei Normal University, Chentow, Hebei, China

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Publications (14)33.65 Total impact

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    ABSTRACT: Adenine phosphoribosyltransferase (APRT) is the key enzyme that converts adenine to adenosine monophosphate (AMP) in the purine salvage pathway. It was found that several different forms of APRT gene exist in plants, but no APRT gene in maize has been reported up to now. In this study, a novel maize APRT gene was cloned and characterized through a combination of bioinformatic, RT-PCR and RACE strategies. The full length of APRT cDNA sequence is 1202 nucleotides, with an ORF encoding 214 amino acid residues. Alignment of the deduced protein with that of other plant APRT genes indicates that the new gene is the form 2 of maize APRT, thus it was named ZmAPT2. Through basic local alignment search tool, search in the genomic survey sequence database of MaizeGDB, the putative genomic sequence of ZmAPT2 was obtained. Comparison of the cDNA and genomic sequence of the ZmAPT2 gene revealed that it contained seven exons and six introns. The locations of the introns within the maize ZmAPT2 coding region were consistent with those in the previously isolated APRTs of arabidopsis and rice. RT-PCR analysis showed that ZmAPRT was constitutively expressing in different organs under high temperature and salt stresses. Southern blot analysis indicated that at least three APRT genes existed in maize genome. These results confirmed that the novel maize ZmAPT2 gene was truly identified, and its potential role in maize growth and development was discussed.
    DNA Sequence 07/2008; 19(3):357-65. · 0.75 Impact Factor
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    ABSTRACT: Southern corn rust (SCR) is a fungal disease caused by Puccinia polysora Underw, which can infect maize and may result in substantial yield losses in maize production. The maize inbred line Qi319 carries the SCR resistance gene RppQ. In order to identify molecular markers linked to the RppQ gene, several techniques were utilized including random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), and amplified fragment length polymorphism (AFLP). In addition, sequence characterized amplified region (SCAR) techniques combined with bulked segregant analysis (BSA) were used. Seven RAPD markers, eight SSR markers, and sixty-three AFLP primer combinations amplified polymorphisms between two parents and two bulk populations. A large F2 population was used for genetic analysis and for fine mapping of the RppQ gene region. One AFLP polymorphic band, M-CAA/E-AGC 324, was converted to a SCAR marker, MA7, which was mapped to a position 0.46 cM from RppQ. Finally, the RppQ gene was mapped between the SCAR marker MA7 and the AFLP marker M-CCG/E-AGA 157 with distances of 0.46 and 1.71 cM, respectively.
    Molecular Genetics and Genomics 01/2008; 278(6):723-8. · 2.88 Impact Factor
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    ABSTRACT: This paper reports the development of SSR markers from EST data and their utilization in germplasm identification of Porphyra. The publicly available EST (expressed sequence tag) sequences of Porphyra were searched from the Internet (http://www.kazura.or.jp/en/plant/porphyra/EST/). From a total of 20,779 obtained EST sequences, 391 SSRs (simple sequence repeats) were analysed with SSRIT software (http://www.gramene.org/db/searches/ssrtool). From those, 48 SSR primer-pairs were designed and tested by commonly used SSR reaction conditions using 22 Porphyra DNA samples as templates. Results showed that 41 SSR primer-pairs gave good amplification patterns. These were used to conduct SSR analyses of genetic diversity and variety identification of the 22 Porphyra lines. A dendrogram and the DNA fingerprints of the Porphyra lines were developed based on the obtained SSR data.
    European Journal of Phycology 08/2006; 41(3):329-336. · 2.34 Impact Factor
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    ABSTRACT: Adenine phosphoribosyl transferase (APRT) is the key enzyme that converts adenine to adenosine monophosphate (AMP) in the purine salvage pathway. A novel form of APRT gene from wheat (Triticum aestivum L.) was cloned through a combination of bioinformatic, RT-PCR and rapid amplification of cDNA ends (RACE) strategies. The total length of the cDNA sequence is 1097 nucleotides, in which an open reading frame (ORF) was found that encodes 221 amino acid residues. Alignment of the deduced amino acid sequence and that of the other plant APRT genes indicates that the novel cDNA in full-length is the form 2 of wheat APRT gene, which was named as TaAPT2. Twelve Single Nucleotide Polymorphism (SNP) sites were identified in the TaAPT2 gene from BNY-S, one wheat temperature-sensitive genic male sterile (TGMS) mutant line derived from the wild type of BNY-F, among which a single base substitution in the purine-binding domain resulted in the Asn to Thr transition. The protein secondary structure prediction revealed that this SNP mutation subsequently resulted in many changes in the quantity and the positions of the helix, extended strand and the loop, which perhaps affect the combining efficiency of the APRT. Northern analysis indicated that the abundance of TaAPT2 gene transcript in the young spikelets reduced obviously in BNY-S under low temperature stress (lower than 10 °C) at the early stage of pollen fertility alternation, however, the transcript of TaAPT2 gene in BNY-F was not affected, indicating that wheat TaAPT2 gene may be related to the fertility alternation and abortion of pollen development.
    Plant Science. 01/2005;
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    ABSTRACT: Twenty-seven Porphyra lines, including lines widely used in China, wild lines and lines introduced to China from abroad in recent years, were screened by random amplified polymorphic DNA (RAPD) technique with 120 operon primers. From the generated RAPD products, 11 bands that showed stable and repeatable RAPD patterns amplified by OPC-04, OPJ-18 and OPX-06, respectively were scored and used to develop the DNA fingerprints of the 27 Porphyra lines. Moreover, the DNA fingerprinting patterns were converted into computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of the corresponding band, respectively. Based on the above results, computerized DNA fingerprints were constructed in which each of the 27 Porphyra lines has its unique fingerprinting pattern and can be easily distinguished from others. Software named PGI (Porphyra germplasm identification) was designed for identification of the 27 Porphyra lines. In addition, seven specific RAPD markers from seven Porphyra lines were identified and two of them were successfully converted into SCAR (sequence characterized amplification region) markers. The developed DNA fingerprinting and specific molecular markers provide useful ways for the identification, classification and resource protection of the Porphyra lines.
    Journal of Applied Phycology 12/2004; 17(1):91-97. · 2.33 Impact Factor
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    ABSTRACT: The moss Physcomitrella patens is an ideal model system for the study of plant physiology and developmental biology. The evolutionary position of moss makes P. patens an ideal subject for the study of land plant evolution. Furthermore, as P. patens is highly tolerant to drought, salt and freezing, it may contain important resistant gene resources. For these reasons, a bacterial artificial chromosome (BAC) library of P. patens was constructed. The high molecular weight (HMW) DNA was partially digested with HindIII. One size fractionation was carried out by pulsed field gel electrophoresis (PFGE) and the selected fragments were ligated to the vector pGUGIBAC1. The ligation mixture was transformed into DH10B cells and recombinants were picked out. In total, 49,920 BAC clones were collected. The BAC library has an average insert size of 65 kb and about seven-fold genome coverage. The hybridization results probed by chloroplast and mitochondrial genes demonstrated that the BAC library has no significant organellar DNA contaminations. The BAC clones were stored using two methods: one involved storing the library in 384-well plates, 1 clone in 1 well; in the other, the library was stored in 96-well plates, 12 clones being pooled together into 1 well. Late embryogenesis-abundant (LEA) proteins are thought to act as a protective component in respond to dehydration, osmotic, and low-temperature stresses. The BAC library was screened with a LEA gene as a probe using rapid PCR methods and six positive clones were screened out. The constructed BAC library will be useful for gene screening and cloning of P. patens. In addition, this BAC library provides a valuable tool for the genomic study of P. patens.
    Plant Science. 01/2004;
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    ABSTRACT: Oryza rufipogon (IRGC105491) is a wild relative of cultivated rice, it contains two favorable yield-enhancing genes (yld1.1 and yld2.1) on chromosomes 1 and 2, respectively, which are capable of improving the yield of hybrid rice by 18 and 17%, respectively. SSR markers RM9, RM24, RM5 and RM306 are flanking yld1.1, while RM166 and RM208 are mapped in the close region to yld2.1. These molecular markers tightly linked to the two yield-enhancing genes were used to screen the plants of backcross population between 9311 (one of the top-performing parental lines in super hybrid rice seed production in China) and O. rufipogon. The results were as follows: (1) in BC2F1 population, the percentage of the individuals which contain both of the O. rufipogon alleles at marker loci RM166 and RM9 was 16.8%; (2) 1.5% individuals of total BC3F1 population have all the six linked markers (RM166, RM9, RM5, RM208, RM24, RM306); (3) in BC4F1 population, the percentage of the individuals which contain both of the two O. rufipogon alleles at marker loci RM166 and RM9 was 18.0%. Based on marker genotypes, the individuals, that contain multiple O. rufipogon markers, were selected and used for further backcross and self cross. Many 9311-type lines with yield-enhancing genes and high yield potential were obtained. After three times self-crossing a stable improved 9311 line was obtained. The results indicated that these molecular markers are feasible for marker-assisted selection (MAS) to screen rice individuals with high yield potential.
    Euphytica 01/2004; 139(2):159-165. · 1.64 Impact Factor
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    ABSTRACT: Maize inbred line 77Ht2 contains agriculturally important genes and has been widely used in corn breeding in China. A bacterial artificial chromosome (BAC) library of 77Ht2 has been constructed in order to identify useful genes and to facilitate the study of the maize genome. The library contains 175104 clones with an average insert size of 57 kb and represents about 4 maize haploid genome equivalents. Characterization of the library showed less than 0.5% of clones to not contain large inserts. Significant contamination of chloroplast and mitochondria DNA was not detected. BAC clones (152 arrays) were stored in 96 microtiter plates, with each well containing 12 clones. This is the first maize BAC library constructed in China. It is well suited for map-based cloning of maize genes and genome physical mapping.
    Plant Molecular Biology Reporter 01/2003; 21(2):159-169. · 5.32 Impact Factor
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    ABSTRACT: Fine mapping of Helminthosporium turcicum resistance gene Ht2 is extremely valuable for map-based cloning of the Ht2 gene, gaining a better knowledge of the distribution of resistance genes in maize genome and marker-assisted selection in maize breeding. An F2 mapping population was developed from a cross between a resistant inbred line 77Ht2 and a susceptible inbred line Huobai. With the aid of RFLP marker analyses, the Ht2 gene was mapped between the RFLP markers UMC89 and BNL2.369 on chromosome 8, with a genetic distance of 0.9 cM to BNL2.369. There was a linkage between SSR markers UMC1202, BNLG1152, UMC1149 and the Ht2 gene by SSR assay. Among the SSR markers, the genetic distance between UMC1149 and the Ht2 gene was 7.2 cM. By bulked segregant analysis 7 RAPD-amplified products which were probably linked to the Ht2 gene were selected after screening 450 RAPD primers and converted the single-copy ones into SCAR markers. Linkage analysis showed that the genetic distance between the SCAR marker SD-06633 and the Ht2 gene was 0.4 cM. From these results, a part of linkage map around the Ht2 gene was constructed.
    Chinese Science Bulletin 01/2003; 48(2):165-169. · 1.37 Impact Factor
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    ABSTRACT: A peach [Prunus persica (L.) Batch] bacterial artificial chromosome (BAC) library of var. Jingyu was constructed. Jingyu is a traditional variety, that displays many of the important agronomic characters of stone fruits. Since peach leaves are rich in polysaccharides, high-molecular-weight (HMW) DNA was extracted from leaf nuclei using a protocol adapted to peach. The HMW DNA embedded in agarose plugs was partially digested by HindIII. After size-selection by pulsed field gel electrophoresis, the selected DNA fragments were ligated to pBeloBAC11 and transformed into E. coli DH10B cells by electroporation. In total 20,736 recombinant clones were obtained. The BAC library has an average insert size of 95 kb and represents approximately 6.7 peach haploid genome equivalents. The BAC clones were stable in E. coli cell after 100 generations. The lack of hybridization to chloroplast and mitochondrial genes demonstrated that the library is predominantly composed of nuclear DNA. The library was screened with two molecular markers, W4 and P20, that are linked to white flesh and nectarine genes of peach, respectively. Ten positive clones were detected. Their fingerprints will be used to determine clone relationships and assemble contigs. This library should be well-suited for the map-based cloning of peach genes and genome physical mapping.
    Theoretical and Applied Genetics 11/2001; 103(8):1174-1179. · 3.66 Impact Factor
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    ABSTRACT: RAPD and RFLP analyses of double haploid lines which derived from hybridization between hexaploid wheat (Triticum aestivum L. 2n=42) and eastern gamagrass (Tripsacum dactyloides L.2n=4x=72) are reported. Two of the 340 Operon primers have been screened, which stably amplifiedTripsacum dactyloides (male parent) specific bands in the double haploid lines. These results confirm the fact thatTripsacum dactyloides DNA has been integrated into wheat genome by sexual hybridization at molecular level. This idea has been further testified by RFLP analysis. Application and potentials of transferringTripsacum dactyloides DNA into wheat genome by sexual hybridization in wheat breeding are discussed.
    Chinese Science Bulletin 03/2000; 45(6):528-532. · 1.37 Impact Factor
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    ABSTRACT: Thermo-sensitive genie male sterile (TGMS) rice has a number of desirable characteristics for hybrid rice production. Many studies have demonstrated that the sterility of TGMS rice is controlled by a single recessive gene. It has been mapped for the first time on chromosome 8 and namedtms 1. Several AFLP markers which tightly linked to thetms 1 gene have been identified recently. In order to develop a detailed physical map of thetms1 gene-encompassing region and finally clone thetms1 gene, a bacterial artificial chromosome (BAC) library of rice 5460F (the fertile mutant line of TGMS rice 5460S) using a modified vector pECBAC1 has been constructed. The constructed 5460F BAC library consists of 16 896 clones with an average insert size of 119 kb, which represents about 4.7 times rice haploid genome equivalents. Neither chloroplast nor mitochondrial DNA was detected from the library. The library was screened with three single copy sequence amplified fragment length polymorphism (AFLP) markers which tightly linked totms1 gene as probes and eight positive clones were identified.
    Chinese Science Bulletin 01/1999; 44(17):1587-1592. · 1.37 Impact Factor
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    ABSTRACT: Thermo-sensitive genie male sterile (TGMS) rice has a number of desirable characteristics for hybrid rice production. Many studies have demonstrated that the sterility of TGMS rice is controlled by a single recessive gene. It has been mapped for the first time on chromosome 8 and namedtms 1. Several AFLP markers which tightly linked to thetms 1 gene have been identified recently. In order to develop a detailed physical map of thetms1 gene-encompassing region and finally clone thetms1 gene, a bacterial artificial chromosome (BAC) library of rice 5460F (the fertile mutant line of TGMS rice 5460S) using a modified vector pECBAC1 has been constructed. The constructed 5460F BAC library consists of 16 896 clones with an average insert size of 119 kb, which represents about 4.7 times rice haploid genome equivalents. Neither chloroplast nor mitochondrial DNA was detected from the library. The library was screened with three single copy sequence amplified fragment length polymorphism (AFLP) markers which tightly linked totms1 gene as probes and eight positive clones were identified.
    Chinese Science Bulletin 01/1999; 44(17):1587-1592. · 1.37 Impact Factor
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    ABSTRACT: Dihydrodipicolinate synthase (DHDPS) is the main enzyme of a specific branch of the aspartate pathway leading to lysine biosynthesis in higher plants. We have cloned and characterized the DHDPS gene from Zizania latifolia Griseb, which was named ZlDHDPS. Sequence analysis indicates that it contains an ORF of 954bp interrupted by two exons and one intron encoding a polypeptide of 317 amino acids lacking a plastid transit peptide and a stop codon. The sequence of ZlDHDPS has high identity with known plant DHDPS in GenBank. Southern blotting analysis indicates that there are two copies of Z. latifolia DHDPS (ZlDHDPS) gene in the Z. latifolia nuclear genome. RT-PCR analysis shows the expression of ZlDHDPS is tissue specific and high level expression is present in fast-growing tissue and reproductive tissue. The 5′-regulatory sequence of ZlDHDPS contains a GT-1 box and a (CA)n element, which may play a role in regulating the expression of ZlDHDPS. The fusion construct of the ZlDHDPS sequence with the reporter gene GUS was transiently expressed in the onion epidermal cells by particle gun-mediated bombardment suggesting that ZlDHDPS was located in the cytoplasm, different from DHDPS gene of other species. Functional complementary analysis showed that ZlDHDPS can recover the DHDPS-deleted mutant of Escherichia coli.
    Plant Molecular Biology Reporter 27(2):199-208. · 5.32 Impact Factor
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    ABSTRACT: New rice lines, restorer line RB207 and maintainer line Yewei B, with better agronomic traits were separately developed from variant progeny of R207 (rice restorer line) and V20B (rice maintainer line) through transformation of genomic DNA ofEchinochloa crusgalli (C4 plant) andOryza minuta, respectively. The phenotypes of the variant lines were apparently different from those of the receptors. Yewei B had stronger tolerance to high temperature than did V20B. The number of spikelets per panicle and the 1000-grain weight of RB207 increased by 40% over those of R207. The results of amplified fragment length polymorphism (AFLP) analysis indicated that the polymorphism rates were both 4.4% between genomes of the variant lines and their receptors. Results demonstrated that special DNA segments fromE. crusgalli andO. minuta might integrate into the genome of cultivated rice and could be stably passed on. The study further shows that transformation of genomic DNA of distant relatives is an effective approach for creating new rice germ plasm.
    Plant Molecular Biology Reporter 22(2):155-164. · 5.32 Impact Factor

Publication Stats

100 Citations
33.65 Total Impact Points

Institutions

  • 2008
    • Hebei Normal University
      Chentow, Hebei, China
  • 2006–2008
    • Northeast Institute of Geography and Agroecology
      • • State Key Laboratory of Plant Genomics
      • • Institute of Genetics and Developmental Biology
      Beijing, Beijing Shi, China
  • 2005
    • Henan Institute of Science and Technology
      Honanfu, Henan Sheng, China
  • 2001–2005
    • Chinese Academy of Sciences
      • Institute of Genetics and Developmental Biology
      Peping, Beijing, China
  • 2004
    • Capital Normal University
      Peping, Beijing, China
    • Agricultural University Of Hebei
      Pao-ting-shih, Hebei, China
  • 1999
    • Technical Institute of Physics and Chemistry
      Peping, Beijing, China