Ward E S, Ellar D J
J Mol Biol. 1986 Sep 5;191(1):1-11. doi: 10.1016/0022-2836(86)90417-1.
The nucleotide sequence of a 1408 base-pair DNA fragment encoding the insecticidal 27,340 Mr delta-endotoxin of Bacillus thuringiensis var. israelensis has been determined by analysis of a recombinant plasmid from Escherichia coli. The hydropathy plot of the protein shows it to be highly hydrophobic, consistent with a postulated cytolytic mechanism of action for the toxin. In addition, the delta-endotoxin transcriptional start points that are used in B. thuringiensis and an E. coli recombinant have been determined. In B. thuringiensis var. israelensis, transcription initiates from a single start point, and gene-specific transcripts are not observed before stage II of sporulation. This is the stage at which delta-endotoxin antigen is first detected, indicating that control of expression is primarily at the transcriptional level for this protein. Analysis of gene-specific transcription in E. coli indicates that at least three start points are utilized in this organism. Interestingly, the highest level of delta-endotoxin mRNA is seen during mid-exponential growth of E. coli and the level appears to decrease as the cells enter the stationary phase of growth.
通过对来自大肠杆菌的重组质粒进行分析,已确定了编码苏云金芽孢杆菌以色列变种杀虫性27340道尔顿δ-内毒素的1408个碱基对DNA片段的核苷酸序列。该蛋白质的亲水性图谱显示其具有高度疏水性,这与推测的毒素溶细胞作用机制一致。此外,还确定了苏云金芽孢杆菌和大肠杆菌重组体中使用的δ-内毒素转录起始点。在苏云金芽孢杆菌以色列变种中,转录从单个起始点开始,在芽孢形成的II期之前未观察到基因特异性转录本。这是首次检测到δ-内毒素抗原的阶段,表明该蛋白质的表达控制主要在转录水平。对大肠杆菌中基因特异性转录的分析表明,该生物体中至少利用了三个起始点。有趣的是,在大肠杆菌指数生长中期观察到δ-内毒素mRNA的最高水平,并且随着细胞进入生长稳定期,该水平似乎会下降。