【作者】 赵娜娜； 艾辉； 王伟娜； 陈冲； 李神斌； 王国秀；

【Author】 Zhao Na-na,Ai Hui,Wang Wei-na,Chen Chong,Li Shen-bin,Wang Guo-xiu (College of Life Sciences,Central China Normal University,Wuhan 430079,China)

【机构】 华中师范大学生命科学学院；

【摘要】 本研究通过RT-PCR同源扩增,首次克隆了棉铃虫(Heliothis armigera)热休克蛋白70(HSP70)cDNA片段。该cDNA片段长1337bp,其中包括开放阅读框(ORF)的起始密码子ATG,推测该序列编码445个氨基酸。序列分析发现,所克隆的棉铃虫HSP70部分序列包含真核生物HSP70高度保守的3个家族标签:IDLGTTYS、IFDLGGGTFDVSIL和IVLVGGSTRIPKVQK。此外,还存在Dank特征基序DLGTT-S-V,ATP/GTP结合位点AEAYLGKT(133-140),非细胞器基序(Non-organellar consensus motif)RARFEEL,核定位序列(Bipartive nuclear localization signals)KRKYKKDLTTNKRALRRL、KRALRRLRTSCERAKRTL和1个糖基化位点NKSI。同源序列比对结果表明,棉铃虫HSP70与其他真核生物的HSP/HSC70有着较高序列同源性(91%—99%),其中与粉纹夜蛾(Trichoplusia ni)热休克蛋白70(HSP70)同源性最高,达99%。系统进化树分析表明,棉铃虫的Hsp70氨基酸残基序列与甘蓝夜蛾(Mamestra brassicae)、粘虫(Mythimna separata)、粉纹夜蛾等物种的Hsp70聚在一支,而与秀丽隐杆线虫(Caenorhabditis elegans)的Hsp70相分离。半定量RT-PCR结果表明,HSP70在对照组和感染组中均有表达,但两者没有显著性差异(P>0.05),初步推断该蛋白是一个结构型HSP70。更多还原

【Abstract】 In this paper,partial cDNA sequence of heat shock protein 70(HSP70) gene of Heliothis armigera was firsdy cloned by using species homology and RT - PCR.The HSP70 cDNA contains 1337 bp,encoding 445 amino acids with an initiator codon ATG.Three highly conserved HSP70 family signatures(IDLGTTYS, IFDLGGGTFDVSIL,IVLVGGSTRIPKVQK) were found in the HSP70 gene.In addition,a putative ATP/GTP binding site(AEAYLGKT),Non - organellar consensus motif(KRKYKKDLTTNKRALRRL),a putative bipartite nuclear location signals(KRKYKKDLTTNKRALRRL and KRALRRLRTSCERAKRTL) and a glycosylation site (NKSI) were detected.Blast analysis revealed that the HSP70 shared high identity(91%—99%) with HSP70s/ HSC70 s of insects,especially with HSP70 from Trichoplusia ni(99%).Phylogenetic tree analysis shows H. armigera is most closely related with Mamestra brassicae,Mythimna separata and T.ni,and isolated from Caenorhabditis elegans.The HSP70 gene expression was examined in both control group and infected group using RT - PCR.The mRNA signal for the gene of the HSP70 was observed in the two groups,and the expression was not significantly different(P>0.5 ).These results suggest that the HSP70 is a structural heat shock protein 70.更多还原