动物医学进展Issue(1)：26-30,5.
微小隐孢子虫类钙调蛋白基因的克隆与原核表达
Cloning and Prokaryotic Expression of C . p arvum Calmodulin-like Protein Gene
摘要
Abstract
In order to express Cryptosporidium parvum calmodulin‐like protein (CML) gene in E .coli BL21(DE3) and analyze the antigenicity of the recombinant protein ,CML gene was amplified by PCR with cDNA of C .parvum oocysts .The amplified CML gene was cloned into pMD18‐T vector and the DNA of recombinant pMD‐CML plasmid was extracted .The plasmid was digested with double enzymes and the ob‐jective fragments were connected with pGEX‐6p‐1 which had been digested with same enzymes .After iden‐tifying by double restrict enzyme digestion and gene sequence analysis , the recombinant plasmids were transformed to E .coli BL21(DE3) cells and the transformed bacteria was induced to express with IPTG . Recombinant proteins were purified by High‐Affinity GST · Bind Resin affinity chromatography .Antigen‐icity of the recombinant proteins was analyzed by Western blot .The results showed that the prokaryotic expression vector pGEX‐CML was constructed successfully and an approximate 51 ku recombinant protein rCML was expressed successfully after inducing with IPTG .The purified recombinant protein could be recognized specifically by the sera from rabbit infected with C .cuniculus .关键词
微小隐孢子虫/类钙调蛋白基因/克隆/原核表达Key words
Cryptosporidium parvum/calmodulin-like protein gene/cloning/prokaryotic expression分类
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杨晓娇,陈兆国,周鹏,米荣升,黄燕,石凯,王晓娟,王向佩,刘宇轩,雷晓思..微小隐孢子虫类钙调蛋白基因的克隆与原核表达[J].动物医学进展,2015,(1):26-30,5.基金项目
国家科技重大专项项目（2012ZX10004220）；中央级公益性科研院所基本科研业务费专项资金项目（2013JB13）；家畜疫病病原生物学国家重点实验室开放基金课题（SKLVEB2013KFKT017）；上海市科技兴农重点攻关项目［沪农科攻字（2005）第3-4号］ （）
