Donovan William P, Engleman James T, Donovan Judith C, Baum James A, Bunkers Greg J, Chi David J, Clinton William P, English Leigh, Heck Gregory R, Ilagan Oliver M, Krasomil-Osterfeld Karina C, Pitkin John W, Roberts James K, Walters Matthew R
Monsanto Company, St. Louis, MO, USA.
Appl Microbiol Biotechnol. 2006 Oct;72(4):713-9. doi: 10.1007/s00253-006-0332-7. Epub 2006 Feb 18.
Bioassay screening of Bacillus thuringiensis culture supernatants identified strain EG2158 as having larvicidal activity against Colorado potato beetle (Leptinotarsa decemlineata) larvae. Ion-exchange fractionation of the EG2158 culture supernatant resulted in the identification of a protein designated Sip1A (secreted insecticidal protein) of approximately 38 kDa having activity against Colorado potato beetle (CPB). An oligonucleotide probe based on the N-terminal sequence of the purified Sip1A protein was used to isolate the sip1A gene. The sequence of the Sip1A protein, as deduced from the sequence of the cloned sip1A gene, contained 367 residues (41,492 Da). Recombinant B. thuringiensis and Escherichia coli harboring cloned sip1A produced Sip1A protein which had insecticidal activity against larvae of CPB, southern corn rootworm (Diabrotica undecimpunctata howardi), and western corn rootworm (Diabrotica virgifera virgifera).
对苏云金芽孢杆菌培养上清液进行生物测定筛选，确定菌株EG2158对科罗拉多马铃薯甲虫（Leptinotarsa decemlineata）幼虫具有杀幼虫活性。对EG2158培养上清液进行离子交换分级分离，鉴定出一种名为Sip1A（分泌型杀虫蛋白）的蛋白质，其分子量约为38 kDa，对科罗拉多马铃薯甲虫（CPB）具有活性。基于纯化的Sip1A蛋白N端序列的寡核苷酸探针被用于分离sip1A基因。从克隆的sip1A基因序列推导的Sip1A蛋白序列包含367个残基（41,492 Da）。携带克隆sip1A的重组苏云金芽孢杆菌和大肠杆菌产生的Sip1A蛋白对CPB、南方玉米根虫（Diabrotica undecimpunctata howardi）和西方玉米根虫（Diabrotica virgifera virgifera）的幼虫具有杀虫活性。
