Chin J Schisto Control ›› 2012, Vol. 24 ›› Issue (6): 663-667.

Previous Articles     Next Articles

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

TANG Jian-xia1|ZHANG Chao1|BAI Liang1|LI Ju-lin1|LIU Kun2|ZHOU Hua-yun1|CAO Jun1|GAO Qi1*   

  1. 1 Jiangsu Institute of Parasitic Diseases|Key Laboratory on Technology for Parasitic Disease Prevention and Control|Ministry of
    Health|Wuxi 214064|China;2 Department of Molecular Microbiology &|Immunology|Johns Hopkins Bloomberg School of Public
    Health|USA
  • Online:2012-12-14 Published:2012-12-19
  • Contact: GAO Qi

中华按蚊水通道蛋白(AsAQP)cDNA克隆与序列分析

唐建霞1|张超1|白亮1|李菊林1|Liu Kun2|周华云1|曹俊1|高琪1*   

  1. 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
  • 通讯作者: 高琪
  • 作者简介:唐建霞|女|博士|助理研究员。研究方向:媒介生物学
  • 基金资助:

    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)

Abstract:

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 cD?
NA of AsAQP consisted of 762 bp and 253 deduced amino acids with a predicted molecular mass of 63.2 kD. Bioinformatics analy?
sis 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. Phy?
logenetic 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 typi?
cal function region. The study lays the foundation for further research on the function of AsAQP.

Key words:  Anopheles sinensis;Aquaporin;Gene cloning;RACE;Sequence analysis

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。

关键词: 中华按蚊;水通道蛋白;基因克隆;RACE;序列分析

CLC Number: