水稻巯基蛋白酶抑制剂基因在烟草叶绿体中的表达  

Expression of Oryzacystatin in Transgenic Tobacco Chloroplast

作　　者：苏宁[1] 冯丽[2] 杨波[1] 孟昆[1] 李轶女[1] 沈桂芳[1] 

机构地区：[1]中国农业科学院生物技术研究所,北京100081 [2]西南农业大学农学系,重庆400716

出　　处：《作物学报》2002年第3期301-304,共4页Acta Agronomica Sinica

基　　金：国家自然科学基金资助 (3 95 70 3 61)

摘　　要：将水稻巯基蛋白酶抑制剂 (Oryzacystatin,OC) c DNA基因克隆、测序分析后 ,与烟草 (N icotiana tabacum L .)叶绿体 16 S r DNA基因启动子和 T393终止子构建表达盒 ,与抗壮观霉素选择标记基因 aad A表达盒相连 ,以烟草叶绿体基因组同源片段 rbc L和 ORF5 12一起构建成叶绿体转化载体 p ZOC。通过基因枪轰击烟草幼苗叶片 ,壮观霉素筛选 ,获得转化再生植株。经 Southern、 Western检测及子代遗传学分析实验证明 OC基因已整合到烟草叶绿体中 ,并且可以遵循非孟德尔的母性遗传规律稳定遗传给后代。After cloning and confirmed by DNA sequencing, Oryzacystatin from rice was placed under the control of 16S rDNA promoter and T 393 terminator from tobacco ( Nicotiana tabacum L.) chloroplast to construct OC expression cassette. It was ligated with selectable marker aad A cassette and homology fragment ( rbc L and ORF 512)of tobacco chloroplast genome to generate transformation vector pZOC. Leaves of tobacco were transformed with particle bombardment method. After screening with spectinomycin, transformants were finally obtained. The analysis of Southern blotting, Western blotting and genetic analysis of the progenies showed that OC gene was introduced into the tobacco chloroplast genome and OC gene was inherited maternally to the progenies. Bioassay showed that the transgenic tobacco did have effective pest resistance to larvae of cotton bollworm ( Helicoverpa zea ) as compared with inferious growth of larvae fed on leaves of untransformed tobacco.

关 键 词：水稻 巯基蛋白酶 抑制剂 基因 烟草 叶绿体 抗虫作用 

分 类 号：S188[农业科学—农业基础科学]