日本蟾蜍聚集素cDNA的克隆与序列分析  

Cloning and Sequence Analysis of Clusterin cDNA of Bufo japonicus formosus

作　　者：袁进强[1] 徐跃[1] 杨仙玉[1] 诸葛慧[1] 张姝芳[1] 

机构地区：[1]浙江农林大学林业与生物技术学院,临安311300

出　　处：《生物技术通报》2013年第1期156-161,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31071181);2011年度浙江省科技创新活动计划项目(2011R412042)

摘　　要：为研究蟾酥、蟾衣、蟾皮中多肽类有效成分,通过菌落PCR对日本蟾蜍(Bufo japonicus formosus)皮肤cDNA质粒文库进行了筛选,获得了聚集素(clusterin,CLU)全长cDNA序列(GenBank登录号为JX035891)并对其进行了生物信息学分析。日本蟾蜍clucDNA全长为1 616 bp,包括1 332 bp的开放阅读框(open reading frame,ORF)、5'端30 bp及3'端254 bp的非翻译区(untranslated region,UTR)。根据cDNA序列推导的日本蟾蜍CLU前体蛋白由443个氨基酸残基组成,其中含20个氨基酸残基组成的信号肽,6个糖基化位点,10个可形成二硫键的半胱氨酸残基和2个卷曲螺旋区域。氨基酸序列同源性分析显示,日本蟾蜍CLU与非洲爪蟾(Xenopus laevis)的同源性为65%,与其他7种动物的同源性则介于41%-48%之间。To study the polypeptide effective components included in Venenum Bufonis, the plasmid eDNA library of adult Japanese toad Bufo japonicus formosus skin was screened. As the result, the clusterin eDNA was obtained （ GenBank accession number ： JX035891 ） . The transcript is 1 616 bp in length containing 30 bp 5＇, 254 bp 3＇ untranslated region （ UTR ） and an open reading frame （ ORF ） of 1 332 bp encoding a polypepfide of 443 amino acids. There is a signal peptide consisted of 20 amino acid residues, six N-glycosylation sites, ten cysteine residues for disulfide bond and two coiled-coil domains in the putative gene product, The homologous analysis indicates the similarity of B. japonicusformosus with Xenopus laevis was 65%, and 41%-48% with other species. The functional characteristics of CLU of B. japonicus formosus seem to be similar as that of human.

关 键 词：日本蟾蜍 聚集素 基因克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]