Ding Xuezhi, Liu Quanlan, Mo Xiangtao, Gao Bida, Xia Liqiu
Department of Microbiology, College of Life Sciences, Human Normal University, Changsha 410081, China.
Wei Sheng Wu Xue Bao. 2003 Jun;43(3):413-7.
In this study, we rapidly identified Bacillus thuringiensis 4.0718 strain that harbored the known cryl and cry2 type genes by a PCR strategy. Three pairs of universal oligonucleotide primers were designed to detect all known cry1, cry2 and cry3 type gene sequences. Then the DNA of the positive strain 4.0718 was probed with a set of specific primers. One feature of this screening method was that each gene was expected to produce a PCR product having a precise molecular weight. PCR products having different sizes probably represented the gene was a potentially novel gene. Differentiations among these genes was determined on the basis of the electrophoresis patterns of PCR products. Finally, five cry1 type genes (cry1 Aa, cry1 Ab, cry1 Ac, cry1 Cb, a novel cry4.5 type genes) and one cry2 Ac type gene had been detected from Bacillus thuringiensis 4.0718 strain.
在本研究中,我们通过PCR策略快速鉴定出携带已知cry1和cry2型基因的苏云金芽孢杆菌4.0718菌株。设计了三对通用寡核苷酸引物来检测所有已知的cry1、cry2和cry3型基因序列。然后用一组特异性引物探测阳性菌株4.0718的DNA。这种筛选方法的一个特点是,每个基因预期会产生具有精确分子量的PCR产物。大小不同的PCR产物可能代表该基因是一个潜在的新基因。这些基因之间的差异是根据PCR产物的电泳图谱确定的。最后,从苏云金芽孢杆菌4.0718菌株中检测到了五个cry1型基因(cry1Aa、cry1Ab、cry1Ac、cry1Cb、一个新的cry4.5型基因)和一个cry2Ac型基因。