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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。转基因植株对棉铃虫幼虫具有抗虫性
After cloned and sequenced the Oryzacystatin (OC) c DNA gene of rice, the expression cassette of 16S rDNA gene promoter and T393 terminator of tobacco chloroplast was constructed and compared with spectinomycin The marker gene aad A expression cassette was ligated. The chloroplast transformation vector p ZOC was constructed with the tobacco chloroplast genome homologous fragment rbc L and ORF5 12. Tobacco seedling leaves were bombarded with gene gun, and spectinomycin was screened to obtain transformed and regenerated plants. The results of Southern and Western analysis and progeny genetic analysis showed that OC gene was integrated into tobacco chloroplast and could be inherited stably to non-Mendelian maternal inheritance. Transgenic plants have insect resistance to cotton bollworm larvae