pcDNA3.1(+)-VEGF165载体的构建及表达蛋白特性分析  

Construction of pcDNA3.1(+)-VEGF165 and Analysis of the Expressed Protein

作　　者：邱明[1] 肖明朝[1] 苟欣[1] 杨钰兴[1] 

机构地区：[1]重庆医科大学附属第一医院泌尿外科,重庆400016

出　　处：《生物技术通报》2010年第2期131-134,161,共5页Biotechnology Bulletin

基　　金：重庆市自然科学基金计划项目(2007BB5288)

摘　　要：构建含人VEGF165基因的重组真核表达质粒,并对其表达蛋白特性进行初步分析。将合成的VEGF165基因序列克隆至pCR2.1-TOPO载体,测序鉴定证实基因碱基序列无误后,再克隆至真核表达载体pcDNA3.1(+),构建成pcD-NA3.1(+)-VEGF165重组质粒。利用DNAstar软件分析基因序列并翻译成氨基酸序列,再用ExPASy protscale和Protean软件分析其疏水特性、蛋白二级结构。经基因测序、酶切鉴定和PCR鉴定证实pcDNA3.1(+)-VEGF165重组质粒构建成功。Ex-PASy protscale和Protean软件分析表明VEGF165蛋白具有较好的水溶性。本研究为VEGF165的基因治疗应用和VEGF165蛋白直接治疗勃起功能障碍奠定了基础。It was to construct the pcDNA3.1 （ ＋ ）-VEGF165 expression vector and analyze the expressed protein. VEGF165 gene was cloned to pCR2.1-TOPO vector and then cloned into the vector pcDNA3.1 （ ＋ ） after sequencing. Two softwares ExPASy protscale and Protean were used to analyze the hydrophobic character and secondary structure of VEGF165 protein. Results indicated that pcDNA3.1 （ ＋ ） -VEGFI65 was successfully constructed and confirmed by gene sequencing and PCR identification. Analysis with ExPASy protscale and Protean showed that the VEGF165 protein has good water-solubility. Indeed,this research provides a foundation for the application of VEGF165 in erectile dysfunction.

关 键 词：血管内皮生长因子 基因克隆 序列测定 基因表达 

分 类 号：Q78[生物学—分子生物学]