摘要
旨在为深入研究Bt毒蛋白的致毒机制和昆虫对Bt毒素的抗性机制提供依据,采用RT-PCR、RACE和qPCR技术克隆并分析暗黑鳃金龟幼虫类钙粘蛋白基因的表达。结果表明,暗黑鳃金龟幼虫类钙粘蛋白基因cDNA序列全长4 833bp(GenBank登录号KM892868),其开放阅读框长4 503bp,编码1 501个氨基酸残基,预测蛋白分子质量169.967ku,理论等电点4.11。在幼虫不同发育时期和3龄幼虫的不同部位均有表达,且在不同时期、不同部位的表达量差异显著。比较不同龄期幼虫的表达量可知,3龄幼虫表达量最高,1龄幼虫表达量最低;比较3龄幼虫不同组织表达量可知,中肠表达量最高,前肠和后肠表达量较低,其他部位则基本不表达。
In order to provide a basis for further research on insecticidal mechanism of Bt in Holotrichia parallela and insect resistances to BT. RT-PCR, RACE and qPCR techniques were used to study the expression of cadherin-like protein in coleoptera Holotrichia parallela larva. The full cDNA sequence （GenBank accession No. KM892868） was 4 833 bp in length,encoding 1 501 amino acid resi dues. The predicted molecular weight and isoelectric point were 169. 967 ku and 4.11 ,respectively. Real-time quantitative PCR showed that CAD-like transcripts were detected in all instars and all body parts of the insect. The expression was significantly different among different instars and different body parts,and it reached the highest level in the third instar and the lowest level in the first instar. Expression peaked in the midgut, while it was almost silent in other body parts except intestine. These results provide a basis for further research on insecticidal mechanism of Bt in H. parallela.
出处
《西北农业学报》
CAS
CSCD
北大核心
2015年第9期152-160,共9页
Acta Agriculturae Boreali-occidentalis Sinica
基金
公益性行业(农业)科研专项(201003025)
关键词
暗黑鳃金龟幼虫
类钙粘蛋白
基因克隆
表达量分析
Holotrichia parallela larvae
Cadherin-like protein
Gene cloning
Expression analysis
