人生长激素基因真核表达载体的构建及鉴定  

Construction and sequence analysis of eukaryotic expression vector for human growth hormone gene

作　　者：杨孔宾[1] 刘炳学[1] 胡恩喜[1] 王宁[1] 刘恩重[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院神经外科,150001

出　　处：《国际遗传学杂志》2010年第2期69-72,共4页International Journal of Genetics

基　　金：国家自然科学基金（30600641）;黑龙江省教育厅基金（11511209）

摘　　要：目的构建人生长激素的真核细胞表达载体，测定并分析目的基因序列。方法用PCR方法扩增出人生长激素基因，连接至T克隆载体，然后分别亚克隆至真核细胞表达载体pCDNA3．1、pCEP4中。使用DNAssist软件分析序列。结果所构建真核表达载体的酶切鉴定、PCR鉴定、测序鉴定均正确。结论成功构建出多个人生长激素基因真核细胞表达载体，验证了基因序列的正确性，为下一步基因治疗研究打下基础。Obiective To construct eukaratic expression vector for human growth hormone gene （hGH） and analyze the sequence for further research work. Methods hGH gene was obtained from pNMG3 with PCR, and ligated into pMD-18-T clone vector, then subcloned into another two expression vector pCDNA3.1 and pCEP4 selectively. The sequence was analyzed with software and compared with the open data in Genebank. Results The constructed vectors were identified with enzyme cutting, PCR, and finally with gene sequencing. The hGH gene of pCDNA3.1-hGH and pCEP4-hGH was the same as pNMG3. However, it was different from the open data of hGH gene in Genebank, and the difference was introns of hGH gene. Conclusion hGH gene in the potent vector - pNMG3 was subcloned into the other two more powerful eukaryotic expression vectors successfully. It is essential for the later gene therapy work to select out an ideal vector for hGH gene.

关 键 词：人生长激素 真核表达载体 基因克隆 

分 类 号：Q78[生物学—分子生物学]