野桑蚕羧酸酯酶基因BmmCarE4和BmmCarE5的克隆及组织表达分析.pdfVIP

野桑蚕羧酸酯酶基因BmmCarE-4 和BmmCarE-5的克隆及组织表达分析 王 东1 李 兵1’2 赵华强1 刘衬丽1 王燕红1 沈卫德1’2 1．苏州大学基础医学与生物科学学院，江苏省苏州市，215123； 2．苏州大学蚕桑研究所，江苏省苏州市，215123 摘 要 为了研究野桑蚕(Bombyx mandarina)与家蚕(Bombyx mori)对杀虫剂抗性差异的 分子机理，采用RT—PCR和RACE方法克隆了野桑蚕羧酸酯酶基因BmmCarE-4的5个选择 性剪切变体序列和BmmCarE-5的两个变体序列。利用半定量RT—PCR方法研究了这两个羧 酸酯酶基因的组织表达特征，结果表明BmmCarE-4和BmmCarE-5具有相似的组织表达特 性。这两个羧酸酯酶基因在中肠中表达量最高，在丝腺和头部表达量次之，在脂肪体和血液中 表达量较低。基因组结构分析表明BmmCarE-4和BmmCarE-5处在同一个转录单元上，而 且共享部分N端外显子序列。氨基酸序列同源性分析表明BmmCarE一4和BmmCarE一5的7 个剪切变体间的氨基酸序列相似性都达52％以上，推测这两个野桑蚕羧酸酯酶基因是由其中 一个基因通过基因加倍复制产生的。 关键词 特种经济动物饲养 野桑蚕羧酸酯酶 组织表达 Cloning and Tissue Expression Analysis of BmmCarE．4 and B Wang Don91 Li Bin91·2 Zhao Huaqian91 Liu Chenlil Wang Yanhon91 Shen Weidel·2 1．SchooI of BasIC MediCine and Unlversi ty，Suzhou 2．IBStItute of SerICUIture，Soochow BiOIogicaI SCIences，Soochow ，dlangsu，215123； UniversIty，Suzhou，diangsu，215123 Abstract In order to research the molecular mechanism that causes insecticide resistance differences between Bombyx mandarina and Bombyx mori，five alternative splicing variants of BmmCarE-4 and 2 variants of BmmCarE-5 were cloned from Bombyx mandarina using RT—PCR and RACE strategies．Semi—quantitative RT—PCR was used to study the tissue ex- pression profiles of the two carboxylesterase genes．The results showed that BmmCarE-4 and 农业现代化与农业新技术 5j3 BmmCarE-5 have similar tissue expression profiles．Their expressions are highest in midgut， a little lower in silk gland and head，and lowest in fat body and hemolymph．Genomic organi— zation analysis indicated that BmmCarE-4 and BmmCarE-5 are located on the same transcrip— tion unit and share partial N—terminal exons．Homology comparison showed that the identi— ties among the 7 alternative splicing variants all reach more than 52％．It was postulated that BmmCarE一4 and BmmCarE-5 were formed through gene duplication of either of the tWO genes． Keywords the rearing of special—type economic animals bombyx mandarina carboxvlesterase tissue expression 野桑蚕(Bombyxmandarina M．)属于鳞翅目蚕蛾科昆虫，和家蚕(Bombyxmori L．)具有