亚洲玉米螟幼虫体内储存蛋白cDNA片段克隆与序列分析  

Cloning and sequence analysis of cDNA fragment of storage protein from the larvae of Asian corn borer,Ostrinia furnacalis(Guenée)(Lepidoptera:Crambidae)

作　　者：冯从经[1] 桑守亮[1] 翟会峰[1] 

机构地区：[1]扬州大学园艺与植物保护学院植物保护系,扬州225009

出　　处：《植物保护学报》2010年第5期391-397,共7页Journal of Plant Protection

基　　金：国家自然科学基金(30971963);扬州大学科技创新培育基金(2010CXJ046)

摘　　要：为研究亚洲玉米螟Ostrinia furnacalis(Guen e)幼虫体内储存蛋白(SP)表达调控的分子机制,根据不同昆虫储存蛋白基因序列的保守区域,设计合成简并引物,采用RT-PCR技术从亚洲玉米螟5龄幼虫中扩增出一段cDNA片段,大小为892bp,编码297个氨基酸,预测分子量为35.7kD,理论等电点pI值为8.73。该基因序列中含有保守的幼虫储存蛋白(LSP)两个特征基序YFTEDIDLNTYYYYFHVDYP和TALRDPVFC。Blast P分析结果表明:该片段推导的氨基酸序列与斜纹夜蛾Spodoptera litura适量甲硫氨酸SP-1的一致性最高,为77%;与小菜蛾Plutella xylostella富含甲硫氨酸SP-1和SP-2、印度谷螟Plodia interpunctellaSP、美国白蛾Hyphantria cuneaSP-2和粉茎螟Sesamia nonagrioides适量甲硫氨酸SP氨基酸序列的一致性分别为74%、74%、72%、71%和65%。构建系统发育树分析了9种鳞翅目昆虫储存蛋白进化关系,结果显示:亚洲玉米螟cDNA片段氨基酸序列与小菜蛾富含甲硫氨酸SP-1的亲缘关系最近,与粉茎螟适量甲硫氨酸SP亲缘关系最远。In order to explore the molecular mechanism of expression and regulation of storage protein（SP） in larvae of Ostrinia furnacalis（Guene）,the sequences of conserved regions of storage protein gene in different insects were used to design the degenerate primers,and a cDNA fragment with the size of 892bp encoding 297 amino acids was amplified from the 5th instar larvae by RT-PCR.The fragment was predicted with molecular weight of 35.7kD and pI value of 8.73.Two conserved larval storage protein（LSP） signature motifs,YFTEDIDLNTYYYYFHVDYP and TALRDPVFC,in the O.furnacalis SP（OfSP） gene were found.Blast P analysis showed that the amino acid sequence of O.furnacalis had the highest identity（77%） to that of Spodoptera litura moderately methionine-rich SP-1,and 74%,74%,72%,71%,65% to the Plutella xylostella methionine-rich SP-1 and SP-2,Plodia interpunctella SP,Hyphantria cunea SP-2,and Sesamia nonagrioides moderately methionine-rich SP,respectively.Phylogenetic analysis of SPs from nine lepidopterans showed that the genetic relationship between OfSP and P.xylostella methionine-rich SP-1 was the closest,while it was the farthest between OfSP and S.nonag-rioides moderately methionine-rich SP.

关 键 词：亚洲玉米螟 储存蛋白 基因克隆 序列分析 

分 类 号：S435.132[农业科学—农业昆虫与害虫防治]