中华按蚊水通道蛋白(AsAQP)cDNA克隆与序列分析  

Cloning and sequence analysis of cDNA encoding aquaporin(AQP) gene from Anopheles sinensis

作　　者：唐建霞 张超[1] 白亮[1] 李菊林[1] Liu Kun 周华云[1] 曹俊[1] 高琪[1] 

机构地区：[1]江苏省血吸虫病防治研究所、卫生部寄生虫病预防与控制技术重点实验室,无锡214064 [2]美国约翰霍普金斯大学分子微生物学与免疫学系

出　　处：《中国血吸虫病防治杂志》2012年第6期663-667,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家自然科学基金(81101276);江苏省自然科学基金(BK2012106、BK2011163);江苏省科教兴卫工程高技术平台(ZX201108)

摘　　要：目的克隆中华按蚊水通道蛋白(AsAQP)基因的cDNA全长序列,分析其基因序列特征,为研究AsAQP的生物学功能提供分子基础。方法根据已报道的昆虫水通道蛋白(AQP)氨基酸序列的保守区域,采用兼并引物从中华按蚊cDNA中获取AsAQP基因片段,在此基础上利用cDNA末端快速扩增(RACE)技术克隆该基因cDNA全长序列,并用生物信息学方法对获取的序列进行分析。结果利用兼并引物从中华按蚊成蚊cDNA中分离到AsAQP基因片段,利用RACE技术克隆到该基因的全长cDNA。序列分析表明,该基因cDNA全长762 bp,编码253个氨基酸,蛋白分子量约为63.2 kD。生物信息学分析表明,AsAQP具有典型的6个跨膜区结构和2个天冬酰胺酸脯氨酸丙氨酸(NPA)结构,该结构是主要内在蛋白(MIP)家族典型的结构特征。AsAQP与致倦库蚊(Culex quinquefasciatus)AQP及埃及伊蚊(Aedes aegypti)AQP蛋白的同源性分别为76%和78%。氨基酸序列聚类分析表明,AsAQP与其他蚊种的水通道蛋白遗传距离较近。结论利用兼并引物结合RACR技术首次获得了编码AsAQP基因的cDNA全长序列,该基因属于MIP蛋白家族成员,具有典型的功能域,为进一步研究该蛋白的功能奠定了基础。Objective To clone and analyze the full-length sequence of aquaporin gene of Anopheles sinensis（AsAQP）,so as to provide an insight into its biology functions.Methods The degenerate primers were used to amplify conserved region of AQP from An.sinensis cDNA.After then,the full-length cDNA of AsAQP was obtained by rapid amplification of cDNA ends（RACE）.Concurrently,the bioinformatics methods were applied to analyze the obtained sequence.Results The obtained full-length cDNA of AsAQP consisted of 762 bp and 253 deduced amino acids with a predicted molecular mass of 63.2 kD.Bioinformatics analysis demonstrated that AsAQP had a typical structure with six membrane-spanning domains and an internal symmetry showing two highly conserved Asn-Pro-Ala（NPA） motif and possessing the consensus sequence of major intrinsic protein（MIP）superfamily.The AsAQP shared the identities of 76% and 78% with those of Culex quinquefasciatus and Aedes aegypti AQPs,respectively.Phylogenetic analysis indicated that AsAQP was clustered with Aedes and Culex AQPs.Conclusions The full-length AsAQP is cloned by degenerate primers and RACE from An.sinensis.The AsAQP gene is a member of MIP protein family,and has the typical function region.The study lays the foundation for further research on the function of AsAQP.

关 键 词：中华按蚊 水通道蛋白 基因克隆 RACE 序列分析 

分 类 号：R382.31[医药卫生—医学寄生虫学]