小鼠α烯醇化酶基因真核表达载体的构建与鉴定  

Construction and Identification of mouse alpha-enolase enzyme gene eukaryotic expression vector

作　　者：贾卫红[1] 罗德炎[2] 段跃强[2] 李志奎[1] 王希良[2] 

机构地区：[1]第四军医大学西京医院呼吸科,西安710032 [2]军事医学科学院微生物流行病研究所免疫室,北京100071

出　　处：《国际免疫学杂志》2011年第5期306-308,共3页International Journal of Immunology

基　　金：国家自然科学基金资助项目

摘　　要：目的构建小鼠α烯醇化酶（En01）基因的真核表达载体。方法以健康小鼠肾脏组织细胞的eDNA为模板，采用聚合酶链式反应（PCR）扩增Enol基因编码区的全部序列，克隆至真核细胞表达载体pCDNA3．1^＋中，测序鉴定目的基因用阳离子脂质体转染的方法转染中国仓鼠卵巢CHO细胞。结果PCR扩增的特异性片段长度为1304bp，以此构建重组质粒pCDNA3．1^＋-Enol，测序结果与Gen．bank中的鼠源Enol基因eDNA序列一致。转染中国仓鼠卵巢细胞后可检测到Enol蛋白的表达。结论构建的真核表达载体pCDNA3．1^＋-Enol可在真核细胞内正确表达，这为进一步的疫苗研究奠定了基础。Objective To construct the eukaryotic expression vector of mouse alpha-enolase enzyme （ Enol ） gene. Methods Enol eDNA of mouse kidney cells was amplified with polymerase chain reaction and inserted into the eukaryotie expression vector of pCDNA3.1^＋. The recombinant plasmid was verified by DNA sequencing and used to transfect Chinese hamster ovary （ CHO ） ceils. Results The amplified fragment was 1 304bp in length. The sequence of Enol gene was in accordance with that in Genbank. The Enol protein was detected in CHO ceils. Conclusion We successfully constructed the pCDNA3.1^＋ -Enol eukaryotic expression vector which may pave a way for further studies in vaccine experiment.

关 键 词：α烯醇化酶 基因克隆 真核表达载体 

分 类 号：Q78[生物学—分子生物学]