兔防御素cDNA表达质粒的构建及其表达  

Construction of Eukaryotic Expression Plasmid of the cDNA of Rabbit Defensins and Its Expression in COS 7

作　　者：张艳宇[1] 刘万臣[1] 张玉静[1] 谭芳[1] 赵建军[1] 柳凤琴[2] 

机构地区：[1]解放军军需大学基础部,吉林长春130062 [2]解放军208医院,吉林长春130062

出　　处：《中国兽医学报》2001年第1期28-30,共3页Chinese Journal of Veterinary Science

基　　金：吉林省青年基金资助项目! (96 35 0 9)

摘　　要：将兔防御素 ( MCP-1 ) c DNA插入真核表达载体 pc DNA3的 Eco R 和 Xba 酶切位点之间 ,构建了兔MCP-1 c DNA的真核表达质粒 pc DEF。通过脂质体转染 ,使兔 MCP-1 c DNA在 COS-7细胞中表达 ,在转染60、84、1 0 8h后 ,提取总 RNA。采用 RT-PCR,在 2 88bp的位置扩增出 1条特异性带 ;RNA斑点印迹杂交表明 ,兔 MCP-1 c DNA在 60、84、1 0 8h均有表达。The eukaryotic expression plasmid pcDEF was constructed by inserting the MCP 1 cDNA sequence into the EcoRⅠ/XbaⅠ fragment of pcDNA3. The recombinant plasmid pcDEF was introduced into the cell of COS 7 by Lipofection. After the introduction of 60 h、84 h and 108 h, the total RNA of COS 7 was extracted and detected by RT PCR. The specific band appearing at the site of 288 bp showed the MCP 1 cDNA expressed at the hour of 60 h, 84 h , 108 h. The PCR assay was utilized to label the probe of Rabbit MCP 1 cDNA. Based on the RT PCR ,the RNA Dot blot hybridization was performed in order to identification the special expression of the Rabbit MCP 1 cDNA .The result of the RNA Dot blot showed that the MCP 1 cDNA expressed after the introduction of 60 h, 84 h and 108 h.

关 键 词：兔防御素 转染 基因表达 斑点印迹杂交 质粒构建 

分 类 号：Q78[生物学—分子生物学] S852.43[农业科学—基础兽医学]