淡色库蚊抗药性相关胰蛋白酶基因cDNA全长克隆与序列分析  

Cloning and Sequence Analysis of Full-length Trypsin cDNA of Culex pipiens pallens

作　　者：沈波[1] 田海生[1] 马磊[1] 李秀兰[1] 郑仕中[1] 周大虎[1] 朱昌亮[1] 

机构地区：[1]南京医科大学寄生虫学教研室,南京210029

出　　处：《生物化学与生物物理学报》2002年第1期28-32,共5页

基　　金：国家自然科学基金资助项目 (No .39970 6 6 6和No .30 170 835 )~~

摘　　要：用逆Northern印迹和Northern印迹法进一步鉴定淡色库蚊对溴氰菊酯抗药性和敏感性品系胰蛋白酶的表达差异 ,结果显示 ,胰蛋白酶基因在抗药性品系中的表达量分别是敏感性品系的 4.3和 3.9倍。采用RACE法筛选cDNA文库 ,获得总长度为 90 9bp的淡色库蚊胰蛋白酶基因的全长序列 ,其中开放阅读框为 786bp ,推导出编码 2 6 1个氨基酸的蛋白质 (GenBank/NCBIAY0 34 0 6 0 ) ,其与冈比亚按蚊胰蛋白酶同源性最高 ,为 5 5 %The expression difference of trypsin gene between deltamethrin-resisitant strain and -susceptible strain of Culex pipiens pallens was further investigated, and the results showed that the expression of trypsin gene were respectively 4.3 and 3.9 fold more in the resistant strain than in the susceptible strain, by reverse Northern blot and Northern blot assays. A full-length trypsin cDNA of 909 base pairs (GenBank/NCBI AY034060) with an open reading frame of 786 base pairs was cloned from the constructed cDNA library by the rapid amplification of cDNA ends, and the deduced protein of 261 amino acids was 55% homologous with Anopheles gambiae trypsin.

关 键 词：淡色库蚊 抗药性 胰蛋白酶基因 逆Northern印迹 NORTHERN印迹 cDNA 全长克隆 序列分析 

分 类 号：Q781[生物学—分子生物学]