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Cloning and analysis of a cDNA encoding acetohydroxy acid isomeroreductase from G2 pea
Authors:Huasong Xu  Yunjian Xu  Xuesong Gu  Tingzhang Hu  Yanhui Su  Yuxian Zhu
Institution:(1) The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, 100871 Beijing, China;(2) College of Biotechnology, South China Agricultural University, 510642 Guangzhou, China;(3) Department of Biology, Sichuan Three-Gorge College, 404000 Chongqing, China
Abstract:Using cDNA representational difference analysis (cDNA RDA) method, we have successfully isolated a gene fragment whose expression was specifically induced by external GA3 application. Screening a G2 pea cDNA library using this fragment as a probe, we obtained a 2036 bp full-length cDNA. It contains a 1746 bp open reading frame and encodes a protein of 581 amino acids with a theoretical molecular weight of 64 ku. It shares high-level sequence identity withAAIR genes from other plant species. This cDNA was cloned into expression vector and recombinantE. coli DH5α cells with remarkable AAIR enzyme activity were obtained.
Keywords:G2 pea  clone  acetohydroxy acid isomeroreductase  prokaryotic expression
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