CAJ | 학술논문

针对转基因水稻 TT51-1插入的 Bt 基因，扩增全长序列并克隆至 pGEM-T 载体．测序验证后利用 HindⅢ与BamHⅠ双酶切将Bt基因定向插入pET-28a载体．转化重组构建的pET-28a-cry至BL21(DE3)宿主菌，在0.1,mmol/L IPTG诱导下获得以包涵体形式表达的目的蛋白．经SDS-PAGE凝胶回收法获得相对分子质量约7.1×104的目的蛋白rCRY，为转基因作物检测新方法的开发提供了物质基础．
침대전기인수도 TT51-1삽입적 Bt 기인，확증전장서렬병극륭지 pGEM-T 재체．측서험증후이용 HindⅢ여BamHⅠ쌍매절장Bt기인정향삽입pET-28a재체．전화중조구건적pET-28a-cry지BL21(DE3)숙주균，재0.1,mmol/L IPTG유도하획득이포함체형식표체적목적단백．경SDS-PAGE응효회수법획득상대분자질량약7.1×104적목적단백rCRY，위전기인작물검측신방법적개발제공료물질기출．
The full-length sequence of integrated Bt gene in genetically modified rice TT51-1,was amplified and cloned into pGEM-T vector. After sequencing was confirmed,the Bt gene was inserted into pET-28a expression vector using HindⅢand BamHⅠ. The recombined pET-28a-cry was transformed into E. coli BL21(DE3)and the recombined target pro-tein(rCRY)was expressed in E. coli BL21(DE3)host cells under 0.1,mmol/L IPTG induction. The purified rCRY protein with the relative molecular mass 7.1×104 was recovered from SDS-PAGE gel by denaturation and refolding of inclusive body protein. This work would be helpful for the development of new GMOs detection methods.