淡色库蚊β-肌动蛋白基因克隆与序列分析  

CLONE AND SEQUENCE ANALYSIS OF β-ACTIN GENE FROM CULEX PIPIENS PALLENS

作　　者：胡莺[1] 沈波[1] 杨明夏[1] 马磊[1] 李秀兰[1] 田海生[1] 朱昌亮[1] 

机构地区：[1]南京医科大学病原生物学系,江苏南京210029

出　　处：《中国寄生虫病防治杂志》2003年第2期65-68,共4页Chinese Journal of Parasitic Disease Control

基　　金：国家自然科学基金资助课题 (No.39970 666和 No.30 1 70 835)

摘　　要：目的　获取淡色库蚊 β-肌动蛋白全长基因。　方法　采用 RACE法筛选淡色库蚊溴氰菊酯抗性品系 c DNA表达文库 ,获得 β-肌动蛋白基因的 5′和 3′RACE序列 ,然后通过 T/ A克隆插入 p GEM- T质粒载体 ,经过测序、拼接获取全长序列。用 BL ASTp软件将该基因推导的氨基酸序列与蛋白质公共数据库 Swissprot比对、分析。　结果　获得淡色库蚊 β-肌动蛋白基因全长序列 ,总长度为 12 90 bp,开放阅读框为 1132 bp,编码 377个氨基酸 (Gen Bank/ NCBIAY10 0 0 0 5 )。推导的氨基酸序列与公共数据库比对 ,发现与冈比亚按蚊和黑腹果蝇的 β-肌动蛋白的同源性均为 91%。蛋白质的理论分子质量为 4 1.7ku,等电点 (PI)为 5 .4。　结论　获得了淡色库蚊 β-肌动蛋白基因全长序列 ,为蚊虫分子生物学的进一步研究奠定了基础。Objective To obtain the sequence of β actin cDNA of Culex pipiens pallens . Methods RACE was used to screen the constructed cDNA library of the resistance strain of Cx. pipiens pallens . The 5′ and 3′ RACE fragments of the β actin cDNA were gotten and cloned into pGEM T vector, then sequenced and spliced. With software BLASTp, the deduced amino acids sequence were aligned to database Swissport. Results A sequence of 1 290 base pairs including an open reading frame of 1 132 base pairs was gotten(GenBank/NCBI AY100005). The deduced protein had 377 amino acids and the theoretic molecular weight and PI were 41.7 ku and 5.4 respectively. Aligned to Swissport, it shared 91% and 91% homology respectively with β actin gene of Anopheles gambiae and Drosophila melanogaster . Conclusion β actin cDNA sequence of Cx. pipiens pallens was gotten. The gene will be used as the internal control in other experiments of Culex mosquitoes.

关 键 词：淡色库蚊 Β-肌动蛋白 克隆 序列分析 cDNA快速末端扩增 

分 类 号：R384.1[医药卫生—医学寄生虫学]