苏云金杆菌新菌株WB9 cry1基因型的PCR-RFLP分析

PCR-RFLP analysis of cry 1-type gene from Bacillus thuringiensis WB9

利用 PCR- RFLP技术对 Bt新菌株 WB9的 cry1基因型进行分析 ,结果表明该菌株含有 cry1Aa、cry1Ab、cry1Cb、cry1Fa和 cry1Ga 5种 cry1基因型 ,且 cry1Ab的酶切产物不同于正常的 cry1Ab.采用特异引物对 G1/ G2扩增 ,也证明 WB9含有 cry1Ab基因 .在此基础上 ,再利用引物对 G1/ K3un2进行扩增 ,其产物经回收纯化后用 Eco R / Xba 双酶切分析 ,结果也表明 WB9所含的 cry1Ab与众不同 ,无大片段出现且有一条 4 0 0 - 5 0 0

PCR RFLP was used to detect the cry 1 type gene from Bt new isolate WB9 and its 5 genes, cry 1Aa, cry 1Ab, cry 1Cb, cry 1Fa and cry 1Ga were identified. However, the RFLP patterns of its cry 1Ab profile were different from the others. With specific cry 1Ab primers, the evidence showed that this isolate also harbours cry 1Ab gene by PCR. A pair of universal primers, which can amplify nearly whole part DNA of cry 1Ab, and 2 restriction enzymes were selected for futher PCR RFLP analysis. Results indicated that the isolate seems to harbour a novel gene.