期刊文献+
共找到198篇文章
< 1 2 10 >
每页显示 20 50 100
Construction of a full bacterial artificial chromosome(BAC) library of Oryza sativa genome 被引量:8
1
作者 TAO QUANZHOU HAIYING ZHAO +1 位作者 LONGFANG QIU GUOFAN HONG.(National Center for Gene Researsh, Chincse Academy of Sciences, Shanghai 200233, China)(Shanghai Institute of Biochemistry, Chinesc Academyof Scienccs, Shanghai 200031, China) 《Cell Research》 SCIE CAS CSCD 1994年第2期127-133,共7页
We have constructed a full BAC library for the superior eaxly indica variety of OryZa sativa, Guang Lu Ai4. The MAX Efficiency DHlOB with increased stabilltyof inserts was used as BAC host cells. The potent pBelo BACI... We have constructed a full BAC library for the superior eaxly indica variety of OryZa sativa, Guang Lu Ai4. The MAX Efficiency DHlOB with increased stabilltyof inserts was used as BAC host cells. The potent pBelo BACII with double selection markers was used as cloning vector. The cloning efficiency we have reached was as high as 98%, and the transformation efficiency was raised up to 1Oo transformants / pg of large fragment DNA. The BAC recombinant transformants were picked at random and analyzed for the size of inserts, which turned out to be of 120 kb in length on average. We have obtained more than 20,000 such BAC clones. According to conventional probabillty equation, they covered the entire rice genome of 420,000 kb in length. The entire length of inserts of the library obtained has the 5- to 6- fold coverage of the genome. To our knowledge, this is the first reported full BAC library for a complex genome. 展开更多
关键词 bacterial artificial chromosome indica Oryza sativa library
下载PDF
Bacterial artificial chromosome library construction of root-knot nematode resistant pepper genotype HDA149 and identification of clones linked to Me3 resistant locus
2
作者 GUO Xiao YANG Xiao-hui +5 位作者 YANG Yu MAO Zhen-chuan LIU Feng MA Wei-qing XIE Bing-yan LI Guang-cun 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第1期57-64,共8页
Pepper (Capsicum annuum. L.) is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode (Meloidogyn... Pepper (Capsicum annuum. L.) is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode (Meloidogyne spp.) is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chro- mosome library (BAC) via the vector of CopyControFM pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90-120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3. 展开更多
关键词 PEPPER bacterial artificial chromosome library.(BAC) root-knot nematode Me3 gene CONTIG
下载PDF
Isolation of Goose-origin scFv Antibodies Against Goose Parvovirus from Bacterial Display Antibody Libraries
3
作者 Zhou Jin-xin Zhang Xiao-yu +5 位作者 Wang Yu-yang Huang Tao Guo Xiao-chen Li De-shan Ma Bo Ren Gui-ping 《Journal of Northeast Agricultural University(English Edition)》 CAS 2021年第1期50-60,共11页
Goose parvovirus(GPV)can cause a highly contagious and fatal gosling plague(GP)disease in goslings and muscoy ducklings.Here,three goose-origin neutralizing single chain variable fragment(scFv)antibodies against GPV S... Goose parvovirus(GPV)can cause a highly contagious and fatal gosling plague(GP)disease in goslings and muscoy ducklings.Here,three goose-origin neutralizing single chain variable fragment(scFv)antibodies against GPV SYG-61 were isolated.The genes of scFv antibodies were derived from goslings immunized with GPV SYG-61,and scFvs were subcloned into a pBSD vector for the construction of pBSD-scFv libraries.The pBSD-scFv libraries were screened following three rounds using VP2(protective antigen of GPV)as the bait by flow cytometry(FCM).After screening,the 15 clones with high mean fluorescence intensity(MFI)were isolated and sequenced.These 15 scFvs were expressed by pET-28a(+)in E.coli.The specificity and affinity of the 15 purified scFvs were successfully confirmed by ELISA.In the preliminary neutralization experiment on primary goose embryo fibroblast(GEF)in vitro,three of the 15 purified scFvs(named scFv-10,scFv-11 and scFv-50)showed significant neutralizing capacities.The study generated the first goose-origin neutralizing scFv against GPV and laid the foundation for the appearance of full-length goose-origin neutralizing monoclonal antibody against GPV. 展开更多
关键词 goose parvovirus(GPV) SCFV bacterial surface display technology antibody library
下载PDF
Comparison of the Bacterial Microbiota in a Bale of Collected Cardboard Determined by 454 Pyrosequencing and Clone Library
4
作者 Valérie Lalande Simon Barnabé Jean-Charles Coté 《Advances in Microbiology》 2014年第12期754-760,共7页
Biofouling, the accumulation of microorganisms, is a major problem in paper mills processing paper and cardboard. This leads to the production of lower quality recycled products. Several studies have focused on the mi... Biofouling, the accumulation of microorganisms, is a major problem in paper mills processing paper and cardboard. This leads to the production of lower quality recycled products. Several studies have focused on the microbial content in the paper mill and the final products. Our aim was to determine the microbial biota in a bale of collected cardboard prior to entering the paper mill. Total genomic DNA was isolated and analyzed using two different methods for comparison purposes: 454 pyrosequencing and clone library. A total of 3268 V6-V8 454 pyrosequencing reads and 322 cloned V6-V8 16S rRNA nucleotide sequences were obtained. Both methods showed the presence of three major bacterial genera: Bacillus, Solibacillus and Paenibacillus, all members of the spore-forming phylum Firmicutes. Pyrosequencing, however, revealed a richer and more diverse bacterial community than clone library. It showed the presence of additional minor Firmicute genera and of a small number of Proteobacteria. The sorting at the recycling plant, the storing, and the processing at the paper mill, the end uses, will all contribute to the bacterial microbiota present in a bale of collected cardboard as revealed here. 展开更多
关键词 454 Pyrosequencing bacterial Microbiota CARDBOARD Clone library Paper Mill
下载PDF
Bacterial diversity in activated sludge from a consecutively aerated submerged membrane bioreactor treating domestic wastewater 被引量:5
5
作者 DU, Cheng WU, Zhenbin +4 位作者 XIAO, Enrong ZHOU, Qiaohong CHENG, Shuiping LIANG, Wei HE, Feng 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2008年第10期1210-1217,共8页
The bacterial diversity of activated sludge from submerged membrane bioreactor (SMBR) was investigated. A 16S rDNA clone library was generated, and 150 clones were screened using restriction fragment length polymorphi... The bacterial diversity of activated sludge from submerged membrane bioreactor (SMBR) was investigated. A 16S rDNA clone library was generated, and 150 clones were screened using restriction fragment length polymorphism (RFLP). Of the screened clones, almost full-length 16S rDNA sequences of 64 clones were sequenced. Phylogenetic tree was constructed with a database containing clone sequences from this study and bacterial rDNA sequences from NCBI for identification purposes. The 90.6% of the clones were a?l... 展开更多
关键词 bacterial diversity restriction fragment length polymorphism (RFLP) submerged membrane bioreactor (SMBR) 16S rDNA clone library
下载PDF
Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance 被引量:1
6
作者 SHEN Wei-wei SONG Cheng-li +3 位作者 CHEN Jie FuYaping WU Jian-li JIANG Shao-mei 《Rice science》 SCIE 2011年第1期17-22,共6页
Based on the sequence of a resistance gene analog FZ14 derived from Zizania latifolia (Griseb.), a pair of specific PCR primers FZ14P1/FZ14P2was designed to isolate candidate disease resistance gene. The pooled-PCR ... Based on the sequence of a resistance gene analog FZ14 derived from Zizania latifolia (Griseb.), a pair of specific PCR primers FZ14P1/FZ14P2was designed to isolate candidate disease resistance gene. The pooled-PCR approach was adopted using the primer pair to screen a genomic transformation-competent artificial chromosome (TAC) library derived from Z. latifolia. A positive TAC clone (ZR1) was obtained and confirmed by sequence analysis. The results indicated that ZR1 consisted of conserved motifs similar to P-loop (kinase la), kinase 2, kinase 3a and GLPL (Gly-Leu-Pro-Leu), suggesting that it could be a portion of NBS-LRR type of resistance gene. Using Agrobacterium-mediated transformation of Nipponbare mature embryo, a total of 48 independent transgenic To plants were obtained. Among them, 36 plants were highly resistant to the virulent bacterial blight strain PXO71. The results indicate that ZR1 contains at least one functional bacterial blight resistance gene. 展开更多
关键词 Zizania latifolia transformation-competent artificial chromosome library resistance-gene analog Oryza sativa bacterial blight resistance gene transfer
下载PDF
Construction of a Three-frame Yeast Two-hybrid cDNA Library of Fusarium oxysporum
7
作者 Luan Fei-shi Li Xiao-mei +1 位作者 Zhu Zi-cheng Wang Xue-zheng 《Journal of Northeast Agricultural University(English Edition)》 CAS 2019年第2期25-32,共8页
A specialized test of two-hybrid library type three-frame cDNA yeast for Muskmelon Fusarium oxysporum using the switching mechanism at the 5'end of RNA template(SMART)technology was constructed to screen for inter... A specialized test of two-hybrid library type three-frame cDNA yeast for Muskmelon Fusarium oxysporum using the switching mechanism at the 5'end of RNA template(SMART)technology was constructed to screen for interaction protein genes for wilt disease and to further research the molecular mechanisms of Fusarium oxysporum pathogenesis to explain the interactions between plant and pathogen.A 500-bp cDNA was purified and extracted using SMART and LD-PCR technology to synthesize ds cDNA and was then homogenized and purified to remove the fragments.After processing,the ds cDNA was connected to three types of reading frame pGADT7-SfiI carriers,and the three connection products in E.coli Electrocell were used to build the primary cDNA library.The titer of three ORF cDNA primary library storage capacities was 2.6×10^6,1.8×10^6 and 3×10^6 cfu;the PCR identification of the ORF 1 and 2 gene recombination rate was 94%,the ORF 3 gene recombination rate was 100%,and the insert length distribution was 0.5-4.0 kb as a single band.To reach the quality requirements for library construction,three kinds of reading frame cDNA primary libraries were mixed and amplified,and the plasmid was transformed into the Y187 yeast strain.The titer of the Y187 yeast library was determined to be 3.5×107 cfu?mL-1,and the base of the yeast library was approximately 1 600 000 cfu.The results showed that the construction of muskmelon Fusarium-specific two-hybrid library type three-frame cDNA yeast had a higher reservoir capacity and recombination rate and met the yeast two-hybrid screening requirements. 展开更多
关键词 MELON FUSARIUM OXYSPORUM YEAST two-hybrid cDNA library normalization
下载PDF
Analysis of Bacterial Communities Present in Agaricus bisporus Phase Ⅱ Compost Using Amplified Ribosomal DNA Restriction Analysis(ARDRA)(Ⅱ)——Analysis of Bacterial Communities
8
作者 HE Lihong YU Rongli +1 位作者 CHEN Mingjie PAN Yingjie 《食用菌学报》 北大核心 2009年第1期6-10,共5页
Changes in the bacterial communities inhabiting Agaricus bisporus compost during Phase Ⅱ treatment were monitored using amplified rDNA restriction analysis(ARDRA).Sequence analysis data indicated that the bacterial c... Changes in the bacterial communities inhabiting Agaricus bisporus compost during Phase Ⅱ treatment were monitored using amplified rDNA restriction analysis(ARDRA).Sequence analysis data indicated that the bacterial communities associated with the compost samples were far richer in composition when determined by ARDRA compared with traditional methods based on bacterial isolation.Furthermore,the composition of the communities,especially in terms of the dominant bacteria during each of the four treatment stages(beginning of Phase Ⅱ,after pasteurization,5 days aeration,end of Phase Ⅱ,respectively),were very different.Restriction endonuclease digestion of mainly bacterial clones from four 16S rDNA libraries,C1,C3,C6 and C7,previously constructed on the basis of 16S rDNA fragments amplified from the four different stages of the Phase Ⅱ composting process,revealed the presence of unique microbial types.Thermophilic bacteria belonging to the Bacilli,and previously unreported strains of Trichococcus,Planococcus,Caryophanon,and subclass γ-Proteobacteria,were identified among the clones from the compost sampled during the aeration period(C6).Bacteria belonging to Thermus thermophilus and subclass α-Proteobacteria were detected in C1 and C7 composts sampled at the beginning and the end of Phase Ⅱ,respectively.Clones of some uncultured bacteria were also scored. 展开更多
关键词 双孢蘑菇 核糖体 细菌克隆 堆肥
下载PDF
Screening proteins that interact with mutant superoxide dismutase 1 from familial amyotrophic lateral sclerosis using a yeast two-hybrid system
9
作者 Guisheng Chen Shugui Shi +7 位作者 Lusi Li Kangning Chen Ju HU Zhenhua Zhou Jun WU GaoxingLuo ShunzongYuan Xu Peng 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第26期2013-2017,共5页
The present study screened a human fetal brain cDNA library to find the proteins that interact with mutant superoxide dismutase 1 (SOD1) using a yeast two-hybrid system. Using BLAST software, 15 real proteins which ... The present study screened a human fetal brain cDNA library to find the proteins that interact with mutant superoxide dismutase 1 (SOD1) using a yeast two-hybrid system. Using BLAST software, 15 real proteins which interacted with mutant SOD1 were obtained, including 8 known proteins (protein tyrosine-phosphatase non-receptor type 2, TBCl D4, protein kinase family, splicing factor, arginine/serine-rich 2, SRC protein tyrosine kinase Fyn, β-sarcoglycan; glycine receptor a2, microtubule associated protein/microtubule affinity-regulating kinase 1, ferritin H chain), and 7 unknown proteins. Results demonstrated interaction of mutant SOD1 with microtubule associated protein/microtubule affinity-regulating kinase 1 and β-sarcoglycan. 展开更多
关键词 yeast two-hybrid system mutant superoxide dismutase 1 cDNA library protein-protein interaction screen amyotrophic lateral sclerosis
下载PDF
Preparation of high molecular weight (HMW) genomic DNA from tea plant (Camellia sinensis) for BAC library construction 被引量:1
10
作者 LIN Jin-ke Dave Kudrna Rod A Wing 《Journal of Agricultural Science and Technology》 2009年第1期1-10,共10页
A bacterial artificial chromosome (BAC) library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight (HMW) genomic DNA is a crucial first step for co... A bacterial artificial chromosome (BAC) library is an invaluable resource tool to initiate tea plant genomics research, and the preparation of high molecular weight (HMW) genomic DNA is a crucial first step for constructing a BAC Library. In order to construct a BAC library for enhancing tea plant genomics research, a new method for the preparation of tea pant high molecular weight (HMW) genomic DNA must be developed due to young tea plant leaves and shoots are notably rich in both tea polyphenols and tea polysaccharides. In this paper, a modified method for preparing high quality tea plant HMW genomi~ DNA was optimized, and the quality of tea plant genomic DNA was evaluated. The results were as follows: Critical indicators of HMW DNA preparation were the appearance of the smooth nuclei in solution (as opposed to sticky-gummy) before agarose plug solidification, non-dark colored nuclei plugs after lysis with an SDS/proteinase K solution, and the quality and quantity of HMW DNA fragments after restriction enzyme digestion. Importantly, 1% dissolved PVP-40 and 1% un-dissolved PVP-40 during the nuclei extraction steps, in conjunction with the removal of PVP-40 from the plug washing and nuclei lysis steps, were critical for achieving HWM tea plant DNA suitable for BAC library construction. Additionally, a third PFGE fraction selection step to eliminate contaminating small DNA fragments. The modifications provided parameters that may have prevented deleterious interactions from tea polyphenols and tea polysaccharides. The HMW genomic DNA produced by this new modified method has been used to successfully construct a large-insert tea plant BAC library, and thus may be suitable for BAC library construction from other plant species that contain similarly interfering compounds. 展开更多
关键词 tea plant bacterial artificial chromosome library BAC clone tea polyphenols high molecular weight genomic DNA preparation Camellia sinensis
下载PDF
Construction and Identification of Bacterial Artificial Chromosome Library for 0-613-2R in Upland Cotton 被引量:4
11
作者 Jian-Mei Yin Wang-Zhen Guo Tian-Zhen Zhang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2006年第2期219-222,共4页
A bacterial artificial chromosome (BAC) library containing a large genomlc DNA insert is an important tool for genome physical mapping, map-based cloning, and genome sequencing. To Isolate genes via a map-based clon... A bacterial artificial chromosome (BAC) library containing a large genomlc DNA insert is an important tool for genome physical mapping, map-based cloning, and genome sequencing. To Isolate genes via a map-based cloning strategy and to perform physical mapping of the cotton genome, a high-quality BAC library containing large cotton DNA Inserts Is needed. We have developed a BAC library of the restoring line 0-613-2R for Isolating the fertility restorer (Rf1) gene and genomic research in cotton (Gossypium hirsutum L.). The BAC library contains 97 825 clones stored In 255 pieces of a 384-well mlcrotiter plate. Random samples of BACs digested with the Notl enzyme Indicated that the average Insert size Is approximately 130 kb, with a range of 80-275 kb, and 95.7% of the BAC clones in the library have an average insert size larger than 100 kb. Based on a cotton genome size of 2 250 Mb, library coverage is 5.7 × haploid genome equivalents. Four clones were selected randomly from the library to determine the stability of the BAC clones. There were no different fingerprints for 0 and 100 generations of each clone digested with Notl and Hlndiii enzymes. Thus, the atabiiity of a single BAC clone can be sustained at iesat for 100 generations. Eight simple sequence repeat (SSR) markers flanking the Rf; gene were chosen to screen the BAC library by pool using PCR method and 25 positive clones were identified with 3.1 positive clones per SSR marker. 展开更多
关键词 bacterial artificial chromosome library COTTON map-based cloning Rf gene.
原文传递
Construction of a Bacterial Artificial Chromosome Library of TM-1,a Standard Line for Genetics and Genomics in Upland Cotton 被引量:3
12
作者 Yan Hu Wang-Zhen Guo Tian-Zhen Zhang 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2009年第1期107-112,共6页
A bacterial artificial chromosome (BAC) library was constructed for Gossypium hirsutum acc. TM-1, a genetic and genomic standard line for Upland cotton. The library consists of 147 456 clones with an average insert ... A bacterial artificial chromosome (BAC) library was constructed for Gossypium hirsutum acc. TM-1, a genetic and genomic standard line for Upland cotton. The library consists of 147 456 clones with an average insert size of 122.8 kb ranging from 97 to 240 kb. About 96.0% of the clones have inserts over 100 kb. Therefore, this library represents theoretically 7.4 haploid genome equivalents based on an AD genome size of 2 425 Mb. Clones were stored in 384 384- well plates and arrayed into multiplex pools for rapid and reliable library screening. BAC screening was carried out by four-round poiymerase chain reactions using 23 simple sequence repeats (SSR) markers, three sequence-related amplified polymorphism markers and one pair of primers for a gene associated with fiber development to test the quality of the library. Correspondingly, in total 92 positive BAC clones were identified with an average four positive clones per SSR marker, ranging from one to eight hits. Additionally, since these SSR markers have been localized to chromosome 12 (A12) and 26 (D12) according to the genetic map, these BAC clones are expected to serve as seeds for the physical mapping of these two homologous chromosomes, sequentially map-based cloning of quantitative trait loci or genes associated with important agronomic traits. 展开更多
关键词 bacterial artificial chromosome library polymerase chain reaction screening simple sequence repeats markers TM-1 upland cotton.
原文传递
Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genic male-sterile rice 5460S 被引量:2
13
作者 邱芳 金德敏 +5 位作者 伏健民 张超良 谢纬武 王斌 杨仁崔 张洪斌 《Science China(Life Sciences)》 SCIE CAS 1999年第6期599-606,共8页
In order to develop a detailed physical map of the thermo-sensitive genie male-sterile (TGMS) gene-encompassing region and finally clone the TGMS gene, a high-quality rice bacterial artificial chromosome (BAC) library... In order to develop a detailed physical map of the thermo-sensitive genie male-sterile (TGMS) gene-encompassing region and finally clone the TGMS gene, a high-quality rice bacterial artificial chromosome (BAC) library from TGMS rice 5460S was constructed. The method of constructing BAC library was examined and optimized. The 5460S library consists of 19 584 BAC clones with an average insert size of 110 kb, which represents about 5 times rice haploid genome equivalents. Rice inserts of up to 140 kb and 250 kb were isolated and appeared stable after 100 generations of serial growth. Hybridization of BAC clones with mitochondria! and chloroplastic genes as probes demonstrated that this library has no organellar contamination. The 5460S library was screened with 3 molecular markers linked to tms 1 gene as probes and at least 1 BAC clone was identified with each probe. The insert ends of positive clones were successfully isolated using thermal asymmetric interlaced PCR (TAIL-PCR) technique. 展开更多
关键词 RICE THERMO-SENSITIVE genie-male STERILE gene bacterial artificial chromosome library thermal asymmetric interlaced PCR.
原文传递
Identification of anti-Mycobacterium tuberculosis agents targeting the interaction of bacterial division proteins FtsZ and SepFe
14
作者 Hongjuan Zhang Ying Chen +6 位作者 Yu Zhang Luyao Qiao Xiangyin Chi Yanxing Han Yuan Lin Shuyi Si Jiandong Jiang 《Acta Pharmaceutica Sinica B》 SCIE CAS CSCD 2023年第5期2056-2070,共15页
Tuberculosis(TB)is one of the deadly diseases caused by Mycobacterium tuberculosis(Mtb),which presents a significant public health challenge.Treatment of TB relies on the combination of several anti-TB drugs to create... Tuberculosis(TB)is one of the deadly diseases caused by Mycobacterium tuberculosis(Mtb),which presents a significant public health challenge.Treatment of TB relies on the combination of several anti-TB drugs to create shorter and safer regimens.Therefore,new anti-TB agents working by different mechanisms are urgently needed.FtsZ,a tubulin-like protein with GTPase activity,forms a dynamic Z-ring in cell division.Most of FtsZ inhibitors are designed to inhibit GTPase activity.In Mtb,the function of Z-ring is modulated by SepF,a FtsZ binding protein.The FtsZ/SepF interaction is essential for FtsZ bundling and localization at the site of division.Here,we established a yeast twohybrid based screening system to identify inhibitors of FtsZ/SepF interaction in M.tuberculosis.Using this system,we found compound T0349 showing strong anti-Mtb activity but with low toxicity to other bacteria strains and mice.Moreover,we have demonstrated that T0349 binds specifically to SepF to block FtsZ/SepF interaction by GST pull-down,fluorescence polarization(FP),surface plasmon resonance(SPR)and CRISPRi knockdown assays.Furthermore,T0349 can inhibit bacterial cell division by inducing filamentation and abnormal septum.Our data demonstrated that FtsZ/SepF interaction is a promising anti-TB drug target for identifying agents with novel mechanisms. 展开更多
关键词 Anti-Mycobacterium tuberculosis FTSZ SepF bacterial division Yeast two-hybrid CRISPRi Protein—protein interaction Inhibitor
原文传递
Use of bacterial two-hybrid system to investigate the molecular interaction between the regulators NifA and NifL of Enterobacter cloacae 被引量:2
15
作者 廖贡献 沈善炯 +1 位作者 俞冠翘 朱家璧 《Science China(Life Sciences)》 SCIE CAS 2002年第6期569-576,共8页
Expression of the nitrogen fixation (nif ) genes is tightly regulated by two proteins NifA and NifL in the (-subdivision of the proteobacteria. NifA is a transcriptional activator, which can be inactivated by NifL in ... Expression of the nitrogen fixation (nif ) genes is tightly regulated by two proteins NifA and NifL in the (-subdivision of the proteobacteria. NifA is a transcriptional activator, which can be inactivated by NifL in the presence of oxygen or excess fixed nitrogen. A direct interaction between E. cloacae NifL and NifA was detected using the bacterial two-hybrid system. This interaction was accelerated in the presence of fixed nitrogen, while oxygen had no effect. NifL proteins, with their C-terminus being deleted, completely lost the ability to interact with NifA. The data suggest that the C-terminal domain of NifL acts as a sensor of the nitrogen status of the cell and mediates interaction with NifA. 展开更多
关键词 bacterial two-hybrid system NifL-NifA complex environmental conditions ENTEROBACTER cloacae Western blotting.
原文传递
Construction of a bacterial artificial chromosome library for Gossypium herbaceum var. africanum 被引量:1
16
作者 GAO HaiYan WANG XingFen +5 位作者 LIU Fang PENG RenHai ZHANG Yan CHENG Hua MA ZhiYing WANG KunBo 《Chinese Science Bulletin》 SCIE EI CAS 2013年第26期3199-3201,共3页
Gossypium herbaceum var. africanum is the only wild cotton species within the cultivated diploid G. herbaceum. As the A sub-genome donor of tetraploid cotton, it is characterized by its resistance to insects, diseases... Gossypium herbaceum var. africanum is the only wild cotton species within the cultivated diploid G. herbaceum. As the A sub-genome donor of tetraploid cotton, it is characterized by its resistance to insects, diseases, and other adversities. We have constructed the first bacterial artificial chromosome library (BAC) for G. herbaceum var. africanum. With high quality and broad coverage, this library includes 75000 clones, with an average insert size of about 115 kb and fewer than 4% empty clones. Our library is approximately five-fold the size of the A-genome (1667 Mb) and it provides 99.3% probability for isolating genes of interest or their sequences. Using nine SSR markers that are located on five different chromosomes and linked with resistance to Verticillium wilt, seven of nine could amplify the 40 superpools and got 1-14 hits. Because of its moderate wide coverage and relative large insert size, this library will be an important genomic resource for classifying and analyzing the evolution of cotton species, as well as for isolating disease-resistance genes and control elements. 展开更多
关键词 细菌人工染色体文库 草棉 变种 非洲 A基因组 棉花品种 SSR标记 基因序列
下载PDF
A study of the protein-protein interactions in the phycocyanin monomer from Synechocystis sp.PCC 6803 using a bacterial two-hybrid system
17
作者 Simi Shao Longsheng Zhao +3 位作者 Pingyi Li Hainan Su Xiulan Chen Yuzhong Zhang 《Engineering Microbiology》 2022年第2期37-41,共5页
Investigations into the intramolecular interactions of the native protein in solution are important to understand its structural stability as well as its potential uses in future applications.In this study,we used a b... Investigations into the intramolecular interactions of the native protein in solution are important to understand its structural stability as well as its potential uses in future applications.In this study,we used a bacterial two-hybrid system to investigate the interaction between the phycocyanin𝛼and𝛽subunits that form the phycocyanin monomer.Key amino acid residues responsible for the interaction between the subunits were identified,providing direct experimental evidence for the intramolecular interaction. 展开更多
关键词 PHYCOCYANIN MONOMER INTERACTION bacterial two-hybrid system
下载PDF
荷斯坦奶牛瘤胃微生物元基因组BAC文库的构建与分析 被引量:27
18
作者 朱雅新 王加启 +4 位作者 马润林 黄力 董志扬 毛爱军 罗淑萍 《微生物学报》 CAS CSCD 北大核心 2007年第2期213-216,共4页
采用未培养技术和脉冲场电泳技术直接从瘤胃微生物提取到大小在2Mb左右混合微生物DNA,经HindⅢ不完全酶切获得50~100kbDNA片段,将其连接在pCC1BAC载体上,转化E.coliEPI300,得到瘤胃微生物BAC文库,经对文库的鉴定分析,该文库的平均插入... 采用未培养技术和脉冲场电泳技术直接从瘤胃微生物提取到大小在2Mb左右混合微生物DNA,经HindⅢ不完全酶切获得50~100kbDNA片段,将其连接在pCC1BAC载体上,转化E.coliEPI300,得到瘤胃微生物BAC文库,经对文库的鉴定分析,该文库的平均插入片段54.5kb,空载体率小于2%,库容837Mb,共保存15360个克隆。通过对该文库进行部分酶活性筛选,获得具有淀粉酶活性的克隆16个;纤维素酶活性的克隆26个,而且能降解纤维素的克隆中25个呈现多酶活性。这些结果表明该文库具有重要研究价值。 展开更多
关键词 瘤胃微生物 细菌人工染色体文库 筛选
下载PDF
同步硝化反硝化系统中反硝化细菌多样性研究 被引量:61
19
作者 郑林雪 李军 +3 位作者 胡家玮 侯爱月 卞伟 郑照明 《中国环境科学》 EI CAS CSCD 北大核心 2015年第1期116-121,共6页
采用聚合酶链式反应(PCR)和分子克隆构建nir S克隆文库对同步硝化反硝化系统好氧池中反硝化细菌多样性进行了研究.从克隆文库中随机挑选75个克隆子进行序列测定,对测序结果进行了BLAST比对.结果表明,有74个克隆子分属于3个不同的细菌类... 采用聚合酶链式反应(PCR)和分子克隆构建nir S克隆文库对同步硝化反硝化系统好氧池中反硝化细菌多样性进行了研究.从克隆文库中随机挑选75个克隆子进行序列测定,对测序结果进行了BLAST比对.结果表明,有74个克隆子分属于3个不同的细菌类群,包括β-Proteobacteria、γ-Proteobacteria和Uncultured bacterium.β-Proteobacteria纲为好氧池内优势菌群,占文库比例的54.41%;其次是γ-Proteobacteria纲,占文库比例的25%.对测序得到的12个OTU用MEGA软件进行系统发育分析,结果显示Thauera属为该系统中最主要的脱氮菌属. 展开更多
关键词 同步硝化反硝化 反硝化细菌 nirS克隆文库 细菌多样性
下载PDF
高覆盖率水稻BAC库的构建及抗病基因相关克隆的筛选 被引量:25
20
作者 王文明 江光怀 +2 位作者 王世全 朱立煌 翟文学 《Acta Genetica Sinica》 SCIE CAS CSCD 北大核心 2001年第2期102-128,共27页
利用含Xa4、xa5和xa13 3个水稻白叶枯病抗性基因的累加系IRBB56构建了一个水稻细菌人工染色体文库。该文库包含55 296个克隆,平均插入片段为132kb。按水稻基因组为450Mb计,该文库覆盖14倍基因组,筛选出任一水稻基因或序列的概率为99.... 利用含Xa4、xa5和xa13 3个水稻白叶枯病抗性基因的累加系IRBB56构建了一个水稻细菌人工染色体文库。该文库包含55 296个克隆,平均插入片段为132kb。按水稻基因组为450Mb计,该文库覆盖14倍基因组,筛选出任一水稻基因或序列的概率为99.99%。用均匀分布的3个叶绿体基因和4个线粒体基因克隆作探针筛选文库,结果显示该文库中含细胞器基因组DNA同源序列的克隆数小于1%。用分布于水稻3条不同染色体、分别与Xa4、xa5和xa13连锁的DNA标记筛选文库,分别检测出11~106个阳性克隆,为克隆这些基因打下了基础。 该文库对水稻基因组的高度覆盖率和较大的插入片段,非常适合于物理作图和基因的分离和克隆。 展开更多
关键词 细菌人工染色体(BAC)文库 基因组 克隆 抗病 水稻 抗病基因
下载PDF
上一页 1 2 10 下一页 到第
使用帮助 返回顶部