Joo Lee Pom, Ahn Ji-Young, Kim Yang-Hoon, Wook Kim Seung, Kim Ji-Yeon, Park Jae-Sung, Lee Jeewon
Department of Chemical and Biological Engineering, Korea University, Anam-Dong 5-1, Sungbuk-Ku, Seoul 136-701, South Korea.
Biochem Biophys Res Commun. 2004 Jul 9;319(4):1110-6. doi: 10.1016/j.bbrc.2004.04.203.
We have identified and cloned a novel toxin gene (tccC1/xptB1) from Xenorhabdus nematophilus strain isolated from Korea-specific entomophagous nematode Steinernema glaseri MK. The DNA sequence of cloned toxin gene (3048 bp) has an open reading frame encoding 1016 amino acids with a predicted molecular mass of 111058 Da. The toxin sequence shares 50-96% identical amino acid residues with the previously reported tccC1 cloned from X. nematophilus, Photorhabdus luminescens W14 P. luminescens TTO1, and Yersinia pestis CO92. The toxin gene was successfully expressed in Escherichia coli, and the recombinant toxin protein caused a rapid cessation in mortality of Galleria mellonella larvae (80% death of larvae within 2 days). Conclusively, the heterologous expression of the novel gene tccC1 cloned into E. coli plasmid vector produced recombinant toxin with high insecticidal activity.
我们从分离自韩国特有的食虫线虫斯氏线虫MK的嗜线虫致病杆菌菌株中鉴定并克隆了一个新的毒素基因(tccC1/xptB1)。克隆的毒素基因的DNA序列(3048 bp)有一个开放阅读框,编码1016个氨基酸,预测分子量为111058 Da。该毒素序列与先前报道的从嗜线虫致病杆菌、发光杆菌W14、发光杆菌TTO1和鼠疫耶尔森菌CO92中克隆的tccC1有50-96%的相同氨基酸残基。该毒素基因在大肠杆菌中成功表达,重组毒素蛋白导致大蜡螟幼虫迅速死亡(2天内80%的幼虫死亡)。总之,克隆到大肠杆菌质粒载体中的新基因tccC1的异源表达产生了具有高杀虫活性的重组毒素。