大鼠pEGFP-C3/BMP-2真核表达载体的构建  

Construction of rat pEGFP-C3/BMP-2 recombinant eukaryotic expressing vector

作　　者：孙欣[1] 曾荣[1] 郭伟韬[1] 肖启贤[1] 王斌[1] 黄云[1] 林颢[1] 

机构地区：[1]广东医学院附属医院骨科,广东湛江524001

出　　处：《中国医药导报》2012年第19期18-20,共3页China Medical Herald

基　　金：广东医学院附属医院青年科研基金(NO.2009k10);湛江市财政资金科技专项竞争性分配项目(湛财工[2009]163号);湛江市科技攻关计划项目(湛科[2011]97号)

摘　　要：目的通过克隆大鼠的BMP2基因,构建EGFP-C3/BMP2基因的真核细胞表达载体。方法把大鼠的基因组DNA通过PCR获得BMP2,克隆构建载体pEGFP/C3-BMP2,并将其转化到大肠杆菌里面,最后进行重组真核表达载体pEGFP-C3-BMP2的构建和鉴定,并可观察其在真核细胞中的表达。结果以大鼠总DNA为模板扩增出1200 bp左右的特异性条带,测序结果与Gene-Bank测序结果相比,翻译成的氨基酸序列相同并完全一致,并可在真核细胞中表达。对重组质粒pEGFP-C3/BMP2进行双酶切鉴定并测序,结果也完全一致。结论为进一步研究利用BMP2基因修饰骨组织工程骨,促进骨折愈合再生提供实验基础。Objective To construct a recombinant eukaryotic expressing vector pEGFP-C3/BMP-2 by using rat bone morphogenetic protein 2 （BMP-2） gene clone. Methods BMP-2 was amplified with PCR and cloned into pEGFP-C3 vector after sequencing, recombinant eukaryotic expressing vector pEGFP-C3/BMP-2 was constructed and identified by sequencing, the expression of BMP-2 in eukaryotic cells was observed and analyzed. Results The sequencing of BMP-2 gene from the rat complied with the Gene-Bank result and with the same amino acid sequence after translation. The recombinant expressing vector pEGFP-C3/BMP-2 was confirmed by double enzyme digestion and sequencing, the successful expression of BMP-2 in eukaryotic cells was observed. Conclusion For the further study BMP2 genetic modification of bone tissue engineering, and promote the regeneration of fracture healing to provide the basis.

关 键 词：BMP2基因 PCR 真核表达载体构建 

分 类 号：R523[医药卫生—内科学]