东亚钳蝎毒液透明质酸酶全基因序列的克隆和结构分析

本研究通过以东亚钳蝎毒腺为研究材料,采用TRIzol法分离透明质酸酶的全基因序列,并从中分离出4个东亚钳蝎透明质酸酶序列,分别命名为BmHYⅠ、BmHYⅡ、BmHYⅢ和BmHYⅣ。BmHYⅠcDNA全长是1423 bp,包括42 bp的5’非翻译区,1230 bp的开放阅读框,编码了一个410个氨基酸,还有151 bp的3’非翻译区;BmHYⅠ中包含5个糖基化位点;与其它物种蛋白质比对结果显示,BmHYⅠ中有10个高度保守半胱氨酸残基形成5个二硫键。BmHYⅠ在二级结构中形成了13个螺旋和14个折叠,其中折叠主要分布在N端和C端。系统进化树结构表明,东亚钳蝎BmHYⅠ与其它蝎子物种透明质酸酶亲缘关系最近,其次是蜘蛛,最后是脊椎动物。而BmHYⅡ、BmHYⅢ和BmHYⅣ序列缺少终止密码子,蛋白序列与BmHYⅠ具有高度同源性。本研究结果为进一步透明质酸酶的功能提供数据参考。

Cloning and Structure Analysis of Scorpion Buthus martensi Venom Hyaluronidases

In this study,the whole gene sequence of hyaluronidase was isolated by TRlzol method with the scorpion venom gland of east Asia as the research material.In the current study,a novel full-length of hyaluronidase BmHYⅠand three noncoding isoforms of BmHYⅡ,BmHYⅢand BmHYⅣwere cloned.A novel hyaluronidase was cloned and named as BmHYⅠ,the full length cDNA was 1423 bp in length containing 42 bp of5’untranslated region(UTR),1230 bp of open reading frame encoded a protein of 410 amino acids,and 151 bp of3’UTR.BmHYⅠhas five potential N-glycosylation sites.The protein alignment result showed that ten conserved Cys resides were well conserved in BmHYⅠcompared with that of others.It showed that 13 helixs and 14 strands are formed in the secondary structure of BmHYⅠ.There was an intensive distribution of the strands in N-terminal and C-terminal.The phylogenetic analysis of sequences provided evidence that BmHYⅠwas closest to the hyaluronidase of scorpion Hottentotta judaicus,then spiders Loxosceles intermedia and Brachypelma vagans,last was verteberate.mRNA sequences of BmHYⅡ,BmHYⅢand BmHYⅣlacked the stop-code in the open reading frame compared with that of BmHYⅠ,but amino acid sequence showed a high-degree homology.The results of this study provide data for further function of hyaluronidase.