浙江枝吻纽虫(Dendrorhynchus zhejiangensis)凝溶胶蛋白和肌动蛋白基因的克隆与序列分析

CLONING AND SEQUENCE ANALYSIS OF GELSOLIN AND ACTIN OF DENDRORHNCHUS ZHEJIANGENSIS

利用已构建的浙江枝吻纽虫cDNA文库,通过PCR技术扩增得到gelsolin和actin基因的全长cDNA序列。其中,gelsolin的cDNA全长1947bp,5′-非翻译区62bp,3′-非翻译区778bp,开放阅读框1107bp,编码369个氨基酸;actin的cDNA全长为1830bp,5′-非翻译区167bp,3′-非翻译区532bp以及1131bp的开放阅读框,编码377个氨基酸。序列分析表明:gelsolin包含3个gelsolin-like(G)结构域,分别含有Ca2+、actin、PIP2结合位点,actin序列中包含了gelsolin、ATP、profiling结合位点。比对结果表明,actin基因氨基酸序列与其它物种的同源性达到94%—99%。

Gelsolin is a versatile actin binding protein,which can regulate actin filament remodeling in a Ca 2＋ -dependant way.In this study,the full length cDNA of gelsolin and actin gene were cloned using the cDNA library of Dendrorhynchus zhejiangensis.The full length cDNA of gelsolin gene was 1947bp with 62bp of 5＇UTR,778bp of 3＇UTR,and 1107bp of ORF,which encodes 369 amino acids.The full length cDNA of actin gene was 1830bp with 167bp of 5＇UTR,532bp of 3＇UTR,and 1131bp of ORF,which encodes 377 amino acids.The analysis of these sequences showed gelsolin gene included three gelsolin-like（G）domains which contained binding sites of Ca 2＋,actin,and PIP2,respectively.Actin included binding sites of gelsolin,ATP,and profiling.The amino acids sequence of actin reached 94%—99% in similarity when aligned with actin proteins of other species.In addition,this study built the foundation for further research on the interaction mechanism between gelsolin and actin,and the mechanism of movement and regeneration of D.zhejiangensis.