CAJ | 학술논문

苏云金芽孢杆菌（Bacillus thuringiensis，Bt）的绝大多数杀虫晶体蛋白（ insecticidal crystal proteins，ICPs）基因是在芽孢形成期(sporulation phase)表达的，而只有少数基因如cry3Aa是在营养期表达的。在本研究中,根据已知的cry3Aa基因启动子序列设计了一对引物（Ep-5s和Ep-3n），利用这对引物从拟步虫甲亚种（Bt subsp. tene brions）中扩增出一个与预期大小（1.1 kb）一致的DNA片段。序列测定及分析结果表明，这个DNA片段含cry3Aa启动子全序列，包括上游AT富含区、两个启动子区、两个SD序列及两组反向重复序列。经过一系列的克隆之后将这个片段克隆到穿梭载体pHT3101上，最后构建成一个B t的营养期表达载体pHPT。
소운금아포간균（Bacillus thuringiensis，Bt）적절대다수살충정 체단백（ insecticidal crystal proteins，ICPs）기인시재아포형성기(sporulation phase)표체적 ，이지유소수기인여cry3Aa시재영양기표체적。재본연구중,근거이지적cry3Aa기 인계동자서렬설 계료일대인물（Ep-5s화Ep-3n），이용저대인물종의보충갑아충（Bt subsp. tene brions） 중확증출일개여예기대소（1.1 kb）일치적DNA편단。서렬측정급분석결과표명，저개DNA편 단함cry3Aa계동자전서렬，포괄상유AT부함구、량개계동자구、량개SD서렬급량조반향 중 복서렬。경과일계렬적극륭지후장저개편단극륭도천사재체pHT3101상，최후구건성일개B t적영양기표체재체pHPT。
The insecticidal activity of Bacillus thuringeinsis (Bt) resides in the parasporal crystalline inclusion body, consisting of a group of insecticidal crystal proteins (ICPs). Most ICP genes are normally sporulation-dependent, wh i le a few of ICP genes such as cry3Aa are non-sporulation-dependent. In thi s stud y, a DNA fragment with the predicated size of 1.1 kb, was obtained by PCR from B t subsp. tenebrions strain with a primer pair (Ep-5s, Ep-3n) designed ac cording to the known sequence of cry3Aa. This DNA fragment contained the completed s eque nce of the cry3Aa promoter, comprising its upstream A+T-rich region, two tr anscr ipts, two Shine-Dalgarno sequences, and two invert-repeats. Through several st ep s of sub-cloning, the promoter was at last inserted into the shuttle vector pHT 3 101 to get a vegetative-expression vector pHPT. And the successful construction of this vector would lay an important foundation for the further research on B t engineering strains.