除草剂抗性基因bar导入烟草叶绿体  

Herbicide Resistance Gene (bar) was Introduced into the Chloroplast of Tobacco

作　　者：张中林[1] 山松[1] 陈曦[1] 刘春英[1] 钱凯先[2] 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 [2]浙江大学生物科学与技术系,浙江杭州310027

出　　处：《作物学报》1999年第5期574-578,共5页Acta Agronomica Sinica

基　　金：本研究系国家科委863专项基金资助

摘　　要：将编码Phosphinothricin acetyltransferase的bar基因与水稻叶绿体psbA基因的启动子和终止子构建成表达盒,连同烟草叶绿体基因组同源片段rpl2-trnH-psbA和trnK-ORF509A以及选择标记基因aadA一起构建成烟草叶绿体转化载体pTZBA.基因枪法转化烟草叶片,经壮观霉素筛选获得转化再生植株.分别以1.0kb的叶绿体同源片段trnK-ORF509A和0.6kb的bar基因为探针,对转基因烟草叶绿体DNA进行Southern杂交检测,证明外源基因已整合入烟草叶绿体基因组中.抗性试验表明,转基因植株具备除草剂(PPT)抗性.The herbicide resistance gene bar encoding Phosphinothricin acetyltransferase was placed under the control of psbA5' and psbA3' of rice to construct bar cassette, which was ligat-ed with the selectable marker aadA gene and these recombined fragments located between the two homology region of tobacco chloroplast genome to generate transformation vector pTZBA. Selection was for spectinomycin resistance after biolistic delivery of plasmid pTZBA DNA into leaf cells. DNA gel-blot analysis probed with homology region trnK-ORF509A and bar confirmed incorporation of the chimeric bar gene into the plastid genome by homologous recombination events via the flanking plastid gene sequences. Some transgenic plants showed resistant to PPT in the test.

关 键 词：BAR基因 同源片段 烟草 aadA基因 除草剂 叶绿体 

分 类 号：S572.01[农业科学—烟草工业] S572.03[农业科学—作物学]