| 链霉菌查尔酮合成酶基因克隆及原核表达载体构建 |
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| 引用本文： | 肖静,吕玉红,李园园,陈雪梅,彭廷文,周春伶,曾令荣,保玉心,凌锌,岳昌武. 链霉菌查尔酮合成酶基因克隆及原核表达载体构建[J]. 重庆文理学院学报, 2013, 32(3): 66-69 |
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| 作者姓名： | 肖静 吕玉红 李园园 陈雪梅 彭廷文 周春伶 曾令荣 保玉心 凌锌 岳昌武 |
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| 作者单位： | 1. 遵义医学院形态学实验室,贵州遵义,563000
2. 遵义医学院贵州省微生物资源及药物开发特色重点实验室,贵州遵义,563000 |
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| 基金项目： | 国家自然科学基金项目，贵州省科学技术基金项目，贵州省科学技术基金项目 |
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| 摘 要： | 根据Genbank数据库已知的链霉菌的查尔酮合成酶基因的保守区设计chs基因特异简并引物,土壤总DNA为模板,利用PCR技术扩增得到该l条chs基因编码区,通过TA克隆、测序和同源比对及进化分析表明:该基因为放线菌来源chs基因.分别在该基因5’末端和3’末端分别引入的限制酶NcoI和EcoR I酶切位点,利用上述2种限制酶分别酶切导入到psimple-T/chs载体和原核表达pET32a,凝胶回收目的片段后,将二者连接并转化大肠杆菌感受态细胞,转化子经菌液PCR筛选、双酶切鉴定后,3730测序结果表明,该基因全长编码区为1089 bp,推测该基因编码全长为362个氨基酸残基,等电点(PI)为5.41、分子量为3 965道尔顿含有CHS保守功能区的酸性蛋白质.分析表明,该基因与Streptomyces lividans来源的查尔酮合成酶RppA基因核苷酸相似性高达93％,氨基酸序列相似性高达87.70％.测序结果表明,该基因已经成功插入到pET32a载体中.
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| 关 键 词： | 查尔酮合成酶 土壤总DNA 基因克隆 原核表达 |
Cloning of streptomycetes chalcone synthase gene and construction of prokaryotic expression carrier |
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| XIAO Jing,LU Yuhong,LI Yuanyuan,CHEN Xuemei,PENG Tingwen,ZHOU Chunling,ZENG Lingrong,BAO Yuxin,LING Xin and YUE Changwu. Cloning of streptomycetes chalcone synthase gene and construction of prokaryotic expression carrier[J]. Journal of Chongqing University of Arts and Sciences, 2013, 32(3): 66-69 |
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| Authors: | XIAO Jing LU Yuhong LI Yuanyuan CHEN Xuemei PENG Tingwen ZHOU Chunling ZENG Lingrong BAO Yuxin LING Xin YUE Changwu |
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| Affiliation: | Morphological Lab of Zunyi Medical College, Zunyi Guizhou 563000, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563000, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563001, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563002, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563003, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563004, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563005, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563006, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563007, China;Microbial Resources and Drug Development Lab of Guizhou Province, Zunyi Medical College, Zunyi Guizhou 563008, China |
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| Abstract: | |
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| Keywords: | Chalcone synthase soil total DNA gene clone prokaryotic expression |
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