灰飞虱Bursicon基因的克隆、序列分析及在不同发育阶段的表达  

Cloning, sequence analysis and developmental expression of the bursicon gene in the small brown planthopper, Laodelphax striatellus (Hemiptera: Delphacidae)

作　　者：杨科[1] 许益鹏[1] 董胜张[1] 俞晓平[1] 

机构地区：[1]中国计量学院生命科学学院,浙江省生物计量及检验检疫技术重点实验室,杭州310018

出　　处：《昆虫学报》2012年第11期1255-1263,共9页Acta Entomologica Sinica

基　　金：国家"973"计划项目(2011CB111602;2012CB114100;2010CB126200);浙江省自然科学基金重点项目(Z3100594);国家"863"计划项目(2012AA021601)

摘　　要：Bursicon是通过G蛋白受体调节昆虫表皮硬化及展翅的功能蛋白,它在昆虫蜕皮后的表皮硬化过程中起着关键作用。为探讨灰飞虱Laodelphax striatellus的bursicon的功能,利用RT-PCR和RACE技术克隆获得1126bp的bursicon α和761bp的bursicon β全长序列,将其分别命名为Lsburs-α和Lsburs-β。生物信息学分析表明:Lsburs-α开放阅读框长483bp,编码160个氨基酸,该蛋白具有2个N-豆蔻酰化位点、3个酪蛋白激酶Ⅱ磷酸化位点以及2个蛋白激酶C磷酸化位点。Lsburs-β开放阅读框长417bp,编码138个氨基酸,该蛋白具有2个N-豆蔻酰化位点、3个酪蛋白激酶Ⅱ磷酸化位点以及1个酪氨酸激酶磷酸化位点。qRT-PCR结果表明:Lsburs-α和Lsburs-β在灰飞虱各龄期均有转录表达,并在若虫期随龄期增加呈上升趋势,在羽化期达到峰值,成虫期表达量逐渐降低。结果提示bursicon与灰飞虱蜕皮后的外表皮硬化关系密切。本文结果为深入研究bursicon的功能、受体调节和信号通路等奠定了基础。Bursicon plays a vital role in insect molting sclerosis, as it regulates cuticle sclerotization （ hardening and tanning） via a G protein-coupled receptor （GPCR）. To study the function of bursicon in the small brown planthopper （SBPH） , Laodelphax striatellus, the full-length sequences of bursicon α and bursicon β were amplified by RT-PCR and RACE, and were designated as Lsburs-α and Lsburs-β, respectively. The products are 1 126 bp and 761 bp in length for Lsburs-α and Lsburs-β, respectively. Lsburs-α contains a 483 bp open reading frame （ORF） encoding a protein with 160 amino acid residues, which has two N-myristoylation sites, three casein kinase Ⅱ phosphorylation sites and two protein kinase C phosphorylation sites. Lsburs-β contains a 417 bp ORF encoding a protein with 138 amino acid residues, which has two N-myristoylation sites, three casein kinase Ⅱ phosphorylation sites and one tyrosine kinase phosphorylation site. Real-time quantitative PCR results showed that the transcripts of Lsburs-α and Lsburs-β were detectable in all the nymphal stages of L. striatellus, increased with the nymphal stages, reached the highest level in the emerging period, and then decreased after adult eclosion. The results suggest that bursicon closely relates to cuticle sclerotization in L. striatellus after molting. The results lay a solid foundation for the further study of bursicon＇ s functions, receptor regulation, signal pathway and so on.

关 键 词：灰飞虱 BURSICON 序列分析 同源性分析 基因表达 实时荧光定量PCR 

分 类 号：Q966[生物学—昆虫学]