Li Ming Shun, Choi Jae Young, Roh Jong Yul, Shim Hee Jin, Kang Joong Nam, Kim Yang-Su, Wang Yong, Yu Zi Niu, Jin Byung Rae, Je Yeon Ho
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Lion Street, Hongshan District, Wuhan 430070, China.
J Microbiol Biotechnol. 2007 Jan;17(1):15-20.
To clone novel cry1-type genes from the Bacillus thuringiensis K1 isolate, about 2.4-kb-long PCR fragments were amplified with two primer sets of ATG1-F/N400-R and 1BeATG1-F/N400-R. Using PCR-RFLP, three novel cry1-type genes, cry1-1, cry1-7, and cry1-44, were obtained from B. thuringiensis K1 and the complete coding sequences of these novel genes were analyzed. The Cry1-1, Cry1-7, and Cry1-44 proteins showed maximum similarities of about 78.0%, 99.7%, and 91.0% with the Cry1Ha1, Cry1Be1, and Cry1Ac2 proteins, respectively. These novel cry1-type genes were expressed using a baculovirus expression vector system and their insecticidal activities were investigated. Whereas all three novel genes were toxic to Plutella xylostella larvae, only Cry1-1 showed insecticidal activity against Spodoptera exigua larvae.
为了从苏云金芽孢杆菌K1分离株中克隆新型cry1类基因,使用两组引物ATG1-F/N400-R和1BeATG1-F/N400-R扩增出约2.4 kb长的PCR片段。利用PCR-RFLP技术,从苏云金芽孢杆菌K1中获得了三个新型cry1类基因cry1-1、cry1-7和cry1-44,并对这些新基因的完整编码序列进行了分析。Cry1-1、Cry1-7和Cry1-44蛋白分别与Cry1Ha1、Cry1Be1和Cry1Ac2蛋白表现出约78.0%、99.7%和91.0%的最大相似性。利用杆状病毒表达载体系统表达了这些新型cry1类基因,并研究了它们的杀虫活性。虽然这三个新基因对小菜蛾幼虫都有毒性,但只有Cry1-1对甜菜夜蛾幼虫表现出杀虫活性。