烟草叶绿体转化载体的构建及转基因植株的获得  

Construction of Vector for Tobacco Plastid Transformation and Obtainment of Tobacco Transgenic Plants

作　　者：邹竹荣[1] 张中林[1] 山松[1] 倪丕冲[1] 沈桂芳[1] 

机构地区：[1]中国农业科学院生物技术研究中心,北京100081

出　　处：《作物学报》1998年第4期410-415,共6页Acta Agronomica Sinica

基　　金：国家自然科学基金;农业部95农-18-03项目

摘　　要：选择烟草叶绿体基因组同源片段rp12-trnH-psbA和trnK-ORF509A,及aadA抗壮观霉素基因,构建烟草叶绿体转化载体pTRZ。制备pTRZ DNA金粉子弹,通过基因枪轰击烟草幼苗叶片,经壮观霉素筛选获得了愈伤组织和转化再生植株。对烟草叶绿体转化植株进行PCR和Southern分析,结果证明其中No.13、16、23为整入了外源aadA的叶绿体转基因植株,同时其子代呈现壮观霉素抗性,aadA基因得到稳定遗传。Vector pTRZ was constructed with rpl2-trnH-psbA, trnK-ORF509A as to bacco plastid homologous recombinant fragments and spectinomycin-resistant aadA gene from plasmid pZS197. After spectinomycin selection, transformed calli and regenerated tobacco plants were obtained from tobacco seedling's leaves bombarded with golden microprojectiles coated with pTRZ DNA. Further PCR and Southern analyses of transformed plants indicated that No. 13, 16, 23 were positive tobacco transplastomic plants and the foreign aadA gene had integrated into their plastid genome. The resistance of progeny phenotype to spectinomycin showed the aadA gene was stably inherited to their progeny.

关 键 词：烟草 叶绿体转化 载体 构建 转基因植株 

分 类 号：S572.03[农业科学—烟草工业]