少棘蜈蚣毒腺RACE cDNA文库的构建及β-actin基因的克隆与序列分析  

Construction of cDNA Library of Venom Gland and Cloning of β-actin cDNA 5′-end Fragments in Scolopendra subspinipes

作　　者：任文华[1] 张双全[1] 宋大祥[1] 周开亚[1] 

机构地区：[1]南京师范大学生命科学学院,南京210097

出　　处：《动物学杂志》2005年第3期1-5,共5页Chinese Journal of Zoology

基　　金：国家自然科学基金资助项目 (No .30 2 70 193)

摘　　要：运用SMART技术首次构建了少棘蜈蚣(Scolopendrasubspinipes)毒腺cDNA文库。经琼脂糖电泳检测,文库所含全长cDNA主要分布在5 0 0bp以上,大于2 0 0 0bp的区域尚有很长拖尾,中间有几条高丰度mRNA亮带。以此文库为模板,通过5′RACE(RapidamplificationofcDNAends,快速扩增cDNA末端)方法获得了细胞质肌动蛋白βactin基因5′末端5 98bp片段,其中包括开放阅读框5 46bp ,编码1 82个氨基酸。将该基因片段推导的氨基酸序列通过BLAST软件与蛋白质公共数据库Swissprot比对,发现与蜜蜂(Apismellifera)的βactin基因同源性高达96% ,说明本实验所构文库质量完全可以满足用RACE方法进行功能基因的cDNA克隆。通过基于双参数模型的NJ法对部分动物βactin基因进行系统重建,较好地反映了这些动物的系统发生关系。A cDNA library of venom gland in Scolopendra subspinipes was constructed with SMART technique. The full length of cDNAs in the library ranged from 500 to 2000 base pairs. A 5′-end of β-actin cDNA sequence of 598 bp in length was obtained with RACE technique, including a 546 bp of open reading frame (ORF) coding 182 amino acids. The BLAST search of the present sequences in the public protein database Swissprot revealled a very high similarity with β-actin gene of Apis mellifera, suggesting that the present library is of high quality for the cDNA cloning of function genes with RACE technique. The phylogenetic reconstruction of some animal β-actin genes based upon NJ algorithm with Kimura two-parameter model could be used to show phylogenetic relationships between different species ofanimals.

关 键 词：cDNA文库 RACE 少棘蜈蚣 n基因 序列分析 毒腺 构建 快速扩增CDNA末端 SMART技术 BLAST软件 全长cDNA cDNA克隆 系统发生关系 开放阅读框 公共数据库 氨基酸序列 基因同源性 双参数模型 电泳检测 mRNA 肌动蛋白 基因片段 

分 类 号：Q785[生物学—分子生物学]