编码日本血吸虫Akt蛋白的cDNA克隆及原核表达  

CLONING AND PROKARYOTIC EXPRESSION OF A CDNA ENCODING AKT PROTEIN OF SCHISTOSOMA JAPONICUM

作　　者：周春景[1] 罗荣[1] 石耀军[1] 丁硕[1,2] 杨德浩[1,3] 程国锋[1] 

机构地区：[1]中国农业科学院上海兽医研究所农业部动物寄生虫学重点开放实验室,上海200241 [2]上海大学,上海200444 [3]河北北方学院,石家庄075000

出　　处：《中国动物传染病学报》2011年第4期62-67,共6页Chinese Journal of Animal Infectious Diseases

基　　金：中央级科研院所基本科研业务费(2008JB18和2009JB08)资助

摘　　要：本研究利用生物信息学分析日本血吸虫编码Akt蛋白的cDNA并对编码该蛋白的cDNA进行了克隆和原核表达。生物信息学分析表明日本血吸虫存在两个Akt蛋白。利用PCR将其中一个Akt蛋白催化功能域的编码cDNA进行了克隆,并利用原核表达系统对此蛋白片段进行诱导表达并进行了重组蛋白的纯化,将重组蛋白免疫新西兰大白兔制备了多克隆抗体。Western blot分析表明,该抗体能被日本血吸虫Akt重组蛋白特异性识别。本研究为进一步研究该蛋白的功能奠定了基础。In the study, we employed bioinformatics and molecular techniques to preliminary study the cDNAs encoding for Akt proteins in Schistosoma japonicum. Bioinformatical analysis indicated that Schistosoma japonicum has two Akt proteins. Subsequently, the cDNA fragment encoding the catalytic domain of a Akt protein was amplified by PCR technique and then cloned into a expression vector pET28a（＋） . The expressed recombinant protein was purified by using a Nickel charged affinity column. The polyclonal antibodies against the Akt protein were raised by immunizing the purified protein in Newland rabbit. Western blot analysis indicated that the polyclonal antibodies were able to specifically recognize the recombinant SjAkt protein. Overall, the present study provides fundamental information for further investigating the functions of Akt proteins in Schistosomajaponicum.

关 键 词：日本血吸虫 原核表达 克隆 信号蛋白 AKT 

分 类 号：S852.735[农业科学—基础兽医学] Q75[农业科学—兽医学]