Ⅱ型胶原蛋白基因cDNA序列分析及其真核表达载体的构建  

Sequence analysis of type Ⅱ collagen protein cDNA and construction of its’eukaryotic expression vector

作　　者：王安宇[1] 魏良纲[1] 乔艺杰[1] 张超[1] 张定安[1] 何光志[2] 

机构地区：[1]贵阳中医学院第二附属医院,贵阳550002 [2]贵阳中医学院基础医学院

出　　处：《山东医药》2012年第28期35-37,共3页Shandong Medical Journal

基　　金：贵州省优秀科技教育人才省长基金项目[黔省专合字(2007)351号];贵州省社会发展科技攻关项目[黔科合SZ字(2009)3047号];2007贵州省教育厅课题;2006贵阳中医学院博士启动基金

摘　　要：目的构建人Ⅱ型胶原蛋白(hCⅡ)基因真核表达载体PcDNA3.1(+)/hCⅡ。方法提取患者关节软骨组织总RNA,应用RT-PCR方法扩增出CⅡ基因片段,测序鉴定正确后,插入真核表达载体PcDNA3.1(+)中构建hCⅡ基因真核表达载体PcDNA3.1(+)/hCⅡ,并进行PCR鉴定、酶切鉴定和序列分析。结果 PCR鉴定、酶切鉴定和序列分析表明CⅡ成功插入表达载体PcDNA3.1(+)。结论本研究成功构建了hCⅡ基因真核表达载体PcDNA3.1(+)/hCⅡ。Objective To construct an eukaryotic expressing vector PcDNA3.1 （ ＋ ）/gene of type Ⅱ collagen （C Ⅱ ） protein. Methods Total RNA from articular cartilage cells in patients with rheumatoid arthritis was extracted and the CⅡ gene fragments were amplified by RT-PCR method. The sequencing was identified, and inserted into the eukaryotic expression vector PcDNA3.1 （ ＋ ） to construct the human C II gene eukaryotic expression vector PcDNA3.1 （ ＋ ）/hC Ⅱ. And construction of expression vector was identified by PCR, restriction endonuclease digestion and sequence analysis. Result C Ⅱ gene was successfully inserted into the expression vector PcDNA3.1 （ ＋ ） by PCR, restriction endonuclease digestion and sequence analysis. Conclusion This study has successfully constructed the eukaryotic expression vector PcD- NA3.1 （ ＋ ）/hC Ⅱ.

关 键 词：Ⅱ型胶原蛋白基因 序列分析 真核表达载体 

分 类 号：R684.3[医药卫生—骨科学]