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Cloning of genomic DNA of rice 5-enolpyruvylshikimate 3-phosphate synthase gene and chromosomal localization of the gene 被引量:3

Cloning of genomic DNA of rice 5-enolpyruvylshikimate 3-phosphate synthase gene and chromosomal localization of the gene
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摘要 The shikimate pathway enzyme 5-enolpyruvylshikimate 3-phosphate synthase (EPSPs) is the target of nonselective herbicide glyphosate. A partial rice epsps cDNA was generated by RT-PCR with primers designed according to EST sequence in GenBank and used as probe for rice genomic library screening. In a screen of approximately 8.0×104 clones from the rice genomic library, sixteen positive clones were obtained, which strongly hybridized to the probe. One clone, E11, was selected for further analysis and the full-length 3661 bp rice epsps genomic sequence was obtained. Sequence analysis and homologous comparison revealed that epsps gene is composed of 8 exons and 7 introns. Analysis by restriction fragment length polymorphism with the probe of rice epsps cDNA fragment confirmed that rice epsps is located on chromosome 6 with an indica-japonica (ZYQ8-JX17) double-haploid (DH) population. This is the first report on the EPSP synthase from monocotyledons. The shikimate pathway enzyme 5-enolpyruvylshikimate 3-phosphate synthase (EPSPs) is the target of nonselective herbicide glyphosate. A partial rice epsps cDNA was generated by RT-PCR with primers designed according to EST sequence in GenBank and used as probe for rice genomic library screening. In a screen of approximately 8.0×104 clones from the rice genomic library, sixteen positive clones were obtained, which strongly hybridized to the probe. One clone, E11, was selected for further analysis and the full-length 3661 bp rice epsps genomic sequence was obtained. Sequence analysis and homologous comparison revealed that epsps gene is composed of 8 exons and 7 introns. Analysis by restriction fragment length polymorphism with the probe of rice epsps cDNA fragment confirmed that rice epsps is located on chromosome 6 with an indica-japonica (ZYQ8-JX17) double-haploid (DH) population. This is the first report on the EPSP synthase from monocotyledons.
出处 《Science China(Life Sciences)》 SCIE CAS 2002年第3期251-259,共9页 中国科学(生命科学英文版)
基金 This work was supported by the National High-Tech Program (863), National Natural Science Foundation of China (Grant Nos. 39989001, 39580012 & 39880023) National Special Program for Research and Industrialization of Transgenic Plants, and Rockefeller F
关键词 5-enolpyruvylshikimate 3-phosphate synthase gene isolation DNA sequence CHROMOSOMAL locating. S-enolpyruvylshikimate 3-phosphate synthase, gene isolation, DNA sequence, Chromosomal locating.
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  • 1P. He,S. G. Li,Q. Qian,Y. Q. Ma,J. Z. Li,W. M. Wang,Y. Chen,L. H. Zhu.Genetic analysis of rice grain quality[J].TAG Theoretical and Applied Genetics (-).1999(3-4)
  • 2S. Ahn,J. A. Anderson,M. E. Sorrells,S. D. Tanksley.Homoeologous relationships of rice, wheat and maize chromosomes[J].MGG Molecular & General Genetics (-).1993(5-6)
  • 3Harry J. Klee,Yvonne M. Muskopf,Charles S. Gasser.Cloning of an Arabidopsis thaliana gene encoding 5-enolpyruvylshikimate-3-phosphate synthase: sequence analysis and manipulation to obtain glyphosate-tolerant plants[J].MGG Molecular & General Genetics.1987(3)
  • 4Liu,Y. G,Shirano,Y,Fakaki,H. et al.Complementation of plant mutants with large genomic DNA fragments by a transformation-competent artificial chromosome vector accelerates positional cloning, Proc.Natl. Acad[].Sci USA.1999

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