Wong H C, Schnepf H E, Whiteley H R
J Biol Chem. 1983 Feb 10;258(3):1960-7.
The nucleotide sequence of the promoter region and part of the coding region of the crystal protein gene from Bacillus thuringiensis var. kurstaki HD-1-Dipel has been determined by analysis of a recombinant plasmid from Escherichia coli. The start points for transcription of the gene in B. thuringiensis and in the E. coli strain carrying the recombinant plasmid were located by S1 nuclease mapping. Two adjacent start sites were identified using RNAs synthesized during sporulation of B. thuringiensis: transcription was initiated from one site early in sporulation and from the other site in the middle of sporulation. A good correlation was found between the appearance of the crystal protein gene-specific RNA and the production of the protein, indicating that the gene is primarily under transcriptional control during sporulation. Parallel studies with the recombinant strain of E. coli revealed the presence of only a single species of gene-specific RNA, regardless of the growth phase of the cells; the crystal protein was produced at all stages of growth. The sequence for eight amino acids at the NH2 terminus of the crystal protein was determined and the corresponding coding sequence was located in the DNA sequence. A potential ribosome binding site of 11 nucleotides was found, located three nucleotides upstream from the initiator ATG codon. The deduced sequence for the first 333 amino acids of the crystal protein is presented.
通过对来自大肠杆菌的重组质粒进行分析，已确定了苏云金芽孢杆菌变种库尔斯塔克HD - 1 - Dipel的晶体蛋白基因启动子区域和部分编码区域的核苷酸序列。利用S1核酸酶图谱定位了该基因在苏云金芽孢杆菌以及携带重组质粒的大肠杆菌菌株中的转录起始点。使用苏云金芽孢杆菌芽孢形成过程中合成的RNA鉴定出两个相邻的起始位点：转录在芽孢形成早期从一个位点起始，在芽孢形成中期从另一个位点起始。在晶体蛋白基因特异性RNA的出现与该蛋白的产生之间发现了良好的相关性，这表明该基因在芽孢形成过程中主要受转录调控。对大肠杆菌重组菌株的平行研究表明，无论细胞处于何种生长阶段，仅存在一种基因特异性RNA；晶体蛋白在生长的各个阶段均有产生。确定了晶体蛋白NH2末端八个氨基酸的序列，并在DNA序列中定位了相应的编码序列。发现了一个11个核苷酸的潜在核糖体结合位点，位于起始ATG密码子上游三个核苷酸处。给出了晶体蛋白前333个氨基酸的推导序列。
