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Generation and Analysis of Pathogenicity-related Gene Mutants of Colletotrichum gloeosporioides Using a Novel Promoter Trapping System 被引量:3
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作者 Zheng Xiaolan Liu Yan +5 位作者 Wei Xiaohui He Chunping Wu Weihuai Liang Yanqiong Li Rui Zheng Fucong 《Plant Diseases and Pests》 CAS 2013年第3期12-15,19,共5页
Agrobactedum tumefac/ens-mediated transformation (ATMT) is an efficient tool for insertional mutagenesis and is used in a wide variety of plants. This paper reports a promoter trapping method to generate mutants in ... Agrobactedum tumefac/ens-mediated transformation (ATMT) is an efficient tool for insertional mutagenesis and is used in a wide variety of plants. This paper reports a promoter trapping method to generate mutants in the filamentous fungus, Colletotrichum gloeosporioides, by ATMT insertion of a trapping vector (pCAHPH) that carries a promoterless hygromycin phosphotransferase (hph) gone. Transformants were selected on the media containing 200 ~mL hy^omycin B, and screened for pathogenicity-related gene mdtants. Their pathogenicity-related mutants T-DNA flanking sequences were then cloned and analyzed. Hph genes were amplified from mutant genomic DNA but not from wild-type DNA, indicating that the phenotypic alternations of these mutants were the results of T-DNA inser- tion. T-DNA flanking sequences were obtained using modified themud asymmetric interlaced PCR. Two right-sided flanking sequences were highly homologous to proteins from other species. 展开更多
关键词 Colletfftrichum gloeosporioides promoter trapping ATMT Pathogenicity-related mutants TaiI-PCR Flanking sequence
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Promoter trapping in Magnaporthe grisea
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作者 刘小红 卢建平 +2 位作者 王教瑜 闵航 林福呈 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2006年第1期28-33,共6页
Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protopl... Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures of M. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis. 展开更多
关键词 promoter trapping Green fluorescent protein Magnaporthe grisea
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OsRRM, a Spen-like rice gene expressed specifically in the endosperm 被引量:2
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作者 Shi-Yan Chen Zong-Yang Wang Xiu-Ling Cai 《Cell Research》 SCIE CAS CSCD 2007年第8期713-721,共9页
We used the promoter trap technique to identify a rice plant, named 107^#, in which the β-glucuronidase (GUS) reporter gene was expressed specifically in the endosperm. A single copy of the T-DNA was inserted into ... We used the promoter trap technique to identify a rice plant, named 107^#, in which the β-glucuronidase (GUS) reporter gene was expressed specifically in the endosperm. A single copy of the T-DNA was inserted into the plant genome, and a candidate gene OsRRM was identified by the insertion. The OsRRM promoter directed GUS expression specifically in rice endosperm, analogous to the GUS expression pattern observed in 107^#. OsRRMis a single-copy gene in rice and encodes a nuclear protein containing 1 005 amino-acid residues with two RNA recognition motifs and one Spen paralog and ortholog C-terminal domain. Westem blot analysis confirmed that the OsRRM protein was specifically expressed in rice endosperm. Ectopic expression of OsRRM in transgenic plants led to abnormalities, such as short stature, retarded growth and low fructification rates. Our data, in conjunction with the reported function of Spen genes, implicated OsRRM in the regulation of cell development in rice endosperm. 展开更多
关键词 RRM Spen promoter trap GUS rice
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Functional genomics: Gene identification via T-DNA mediated gene trap tagging in plants 被引量:2
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作者 唐巍 《Journal of Forestry Research》 SCIE CAS CSCD 2001年第1期1-6,共9页
The fully sequenced genomes of Arabidopsis, rice, tomato, potato, ma ize, wheat, and soybean offer large amounts of information about cellular and de velopmental biology. It is a central challenge of genomics to use t... The fully sequenced genomes of Arabidopsis, rice, tomato, potato, ma ize, wheat, and soybean offer large amounts of information about cellular and de velopmental biology. It is a central challenge of genomics to use this informati on in discovering the function of proteins and identifying developmentally impor tant genes. Although classical genetic approaches to gene identification which r ely on disruption of a gene leading to a recognizable phenotype continues to be an extremely successful one, T-DNA mediated gene trap tagging which has been dev eloped that utilize random integration of reporter gene constructs has also prov en to be an extremely powerful tool in plant cellular developmental biology. In this review, how gene trap tagging, promoter trap tagging, and enhancer trap tag ging detection systems have been applied to plant biology is described and these gene identification techniques could be useful to the plant molecular biology a nd plant biotechnology community. 展开更多
关键词 GENOMICS Gene identification Enhancer trap promoter trap Gene trap
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Molecular analysis of rice plants harboring a multi-functional T-DNA tagging system 被引量:5
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作者 Yimian Ma Luo Liu +10 位作者 Chengguang Zhu Changhui Sun Bo Xu Jun Fang Jiuyou Tang Anding Luo Shouyun Cao Gupo Li Qian Qian Yongbiao Xue Chengcai Chu 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2009年第5期267-276,共10页
About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants... About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants were single copy integration and the average T-DNA insertion number was 2.28. By extensive phenotyping in the field, quite a number of agronomically important mutants were obtained. Histochemical GUS assay with 4,310 primary mutants revealed that the GUS-staining frequency was higher than that of the previous reports in various tissues and especially high in flowers. The T-DNA flanking sequences of some mutants were isolated and the T-DNA insertion sites were mapped to the rice genome. The flanking sequence analysis demonstrated the different integration pattern of the right border and left border into rice genome. Compared with Arabidopsis and poplar, it is much varied in the T-DNA border junctions in rice. 展开更多
关键词 rice mutant population promoter trap activation tag T-DNA integration
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