细粒棘球蚴SmadA基因的克隆及其功能的初步鉴定

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (12): 1756-1762.doi:

细粒棘球蚴SmadA基因的克隆及其功能的初步鉴定

李静1,2 ，张传山2 ，吕国栋2，王俊华2，魏绪法2，林仁勇2，剡根强1*

1. 石河子大学动物科技学院，石河子 832003；2. 新疆医科大学第一附属医院医学研究中心/新疆包虫病基础医学重点实验室，乌鲁木齐 830054

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-12-15 发布日期:2011-12-15
通讯作者: 剡根强

Cloning and Preliminary Characterization of a SmadA Gene from Echinococcus granulosus

LI Jing1,2 , ZHANG Chuanshan2, LV Guodong2, WANG Junhua2,
WEI Xufa2, LIN Renyong2, YAN Genqiang1*

1. College of Animal Science &Technology, Shihezi University, Shihezi 832003, China; 2. Medical Research Center and Xinjiang Key Laboratory of Echinococcosis, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-12-15 Published:2011-12-15
Contact: YAN Genqiang

摘要/Abstract

摘要： 本研究旨在克隆细粒棘球蚴Smad蛋白家族基因EgSmadA，鉴定其结构及表达部位，并建立酵母双杂交体系。利用生物信息学方法, 克隆及分析EgSmadA基因的DNA全长序列，采用免疫组化技术鉴定EgSmadA蛋白的表达及分布，并构建该基因酵母双杂交载体。结果表明，成功扩增出EgSmadA基因组序列全长4 069 bp，包含4个外显子和3个内含子，开放阅读框为957 bp，编码318个氨基酸，该蛋白C端含有Smad蛋白家族进化高度保守的典型结构域MH2 (Mad homology)，并成功构建该基因及其MH2结构域的酵母双杂交载体。免疫组化定位该蛋白主要表达于原头蚴被膜及囊泡生发层。研究结果为进一步研究EgSmadA在细粒棘球蚴生长发育中的作用奠定了基础。

Abstract: The objectives of this study were to clone the EgSmadA gene of Smad family from Echinococcus granulosus, identify its structure and expression profile of the protein, and construct its yeast twohybrid system. The EgSmadA gene was cloned and analyzed by bioinformatics, and detected its expression profile by immunohistochemisty. The results showed that EgSmadA genomic DNA is 4 069 bp, contains 4 exons, 3 introns and a complete open reading frame of 957 bp in length, which encodes for a peptide of 318 amino acids. A conserved MH2 (Mad homology) domain of Smad family was detected in the Cterminal of EgSmadA, and the yeast twohybrid system of EgSmadA and EgSmadAMH2 were successfully constructed. Immunolocalization results indicated that EgSmadA was expressed in the tegument of protoscolex and cellular germinal layer of metacestode. The results provided some new clues for further studying the biological functions of EgSmadA in the growth and development of Echinococcus.

李静;;张传山;吕国栋;王俊华;魏绪法;林仁勇;剡根强. 细粒棘球蚴SmadA基因的克隆及其功能的初步鉴定[J]. 畜牧兽医学报, 2011, 42(12): 1756-1762.

LI Jing;;ZHANG Chuanshan;LV Guodong;WANG Junhua; WEI Xufa;LIN Renyong;YAN Genqiang . Cloning and Preliminary Characterization of a SmadA Gene from Echinococcus granulosus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(12): 1756-1762.

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