单 位：
云南农业大学园林园艺学院;云南省农业生物技术重点实验室
关键词：
Cry1Ab基因;烟草;叶绿体转化;转基因植株
摘 要：
构建了苏云金杆菌晶体毒蛋白基因Cry1Ab的烟草叶绿体表达载体pCTBt,该载体以烟草叶绿体基因组trnI-trnA为同源重组片段,含有壮观霉素抗性筛选标记基因aadA、质体特异性启动子Prrn和终止子TpsbA.利用基因枪转化法,将pCTBt导入栽培烟草红花大金元叶绿体,经过4次继代筛选,获得9个抗壮观霉素的转化株系,转化频率为0.3～0.6株/枪,其中有5个株系PCR检测结果为阳性,初步确定Cry1Ab基因已整合到烟草叶绿体中.转基因烟草抗斜纹夜蛾试验表明,幼虫死亡率为10%～75%,转基因烟草植株具有较好的杀虫活性.
译 名：
Integration of Cry1Ab gene into chloroplast genome of tobacco and generation of transformed plants
作 者：
DING Yu-mei1,YANG Zheng-an2,Zhou Xiao-gan1,ZHANG Shao-song1,SUN Mao-lin1(1.Key Laboratory of Agricultural Biotechnology of Yunnan,Kunming 650223,China;2.College of Horticulture and Landscape,Yunnan Agricultural University,Kunming 650201,China)
关键词：
Cry1Ab gene;nicotiana tabacum;chloroplast transformation;transgenic plant
摘 要：
Chloroplast transformation has become an attractive alternative technology in higher plants.Here,we constructed an expression vector pCTBt used in chloroplast transformation of tobacco,which contains Bacillus crystal toxin Cry1Ab gene as an functional gene,trnI-trnA gene fragment from tobacco plastome as homologous targeting sequences,and the selectable marker gene aadA under the control of a strong plastid promoter Prrn and terminator TpsbA.The vector pCTBt was then bombarded into leaf explants of tobacco cultivar 'Red Flower Dajinyuan' using PDS-1000/He biolistic particle delivery system.A total of 9 transgenic lines resistance to spectinomycin were obtained out of 20 plates bombarded with pCTBt after 16 weeks selection culture.The transformation efficiency(about one recovering plant from 2 or 3 bombardments) was lower compared to that of Maliga's reports(one plant per bombardments).The PCR analyses showed that 5 out of 9 lines performed positive,thus confirmed that the Cry1Ab gene were integrated into the chloroplast genome of tobacco.The bioassays showed the transgenic tobacco lines had a considerable resistance to the larvae of Prodenia litura at the lethal rate of 10% to 75%.
