Shibano Y, Yamagata A, Nakamura N, Iizuka T, Sugisaki H, Takanami M
Gene. 1985;34(2-3):243-51. doi: 10.1016/0378-1119(85)90133-7.
The insecticidal crystal protein (ICP) gene, icp, from a 68-kb plasmid derived from Bacillus thuringiensis subsp. sotto was cloned in Escherichia coli. The icp expression in E. coli cells was confirmed by both immunological and insect-toxicity assays of the cell extract. The entire icp gene resides in the 6.6-kb PstI fragment, which codes for a 144-kDal peptide identical to the intact ICP, as determined by its size and reaction with anti-ICP antibody. Deletion analysis further revealed that the 2.8-kb region within the 6.6-kb PstI fragment codes for ICP. Analysis of the nucleotide sequence indicated that a peptide of 934 amino acid residues truncated at the C-terminal end is encoded by this 2.8-kb fragment. A unique feature of this truncated ICP is the abundance of cysteine and lysine residues within its C-terminal region.
从苏云金芽孢杆菌亚种sotto的一个68 kb质粒中分离出的杀虫晶体蛋白(ICP)基因icp,被克隆到大肠杆菌中。通过对细胞提取物进行免疫和昆虫毒性测定,证实了icp在大肠杆菌细胞中的表达。整个icp基因位于6.6 kb的PstI片段中,根据其大小和与抗ICP抗体的反应确定,该片段编码一个与完整ICP相同的144 kDal肽。缺失分析进一步表明,6.6 kb PstI片段内的2.8 kb区域编码ICP。核苷酸序列分析表明,这个2.8 kb片段编码一个在C末端截短的934个氨基酸残基的肽。这种截短的ICP的一个独特特征是其C末端区域富含半胱氨酸和赖氨酸残基。