CAJ | 학술논문

目的构建小鼠α烯醇化酶(Enol)基因的真核表达载体。方法以健康小鼠肾脏组织细胞的cDNA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1 304bp，以此构建重组质粒pCDNA3.1+-Enol，测序结果与Genbank中的鼠源Enol基因cDNA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。
목적구건소서α희순화매(Enol)기인적진핵표체재체。방법이건강소서신장조직세포적cDNA위모판，채용취합매련식반응(PCR)확증Enol기인편마구적전부서렬，극륭지진핵세포표체재체pCDNA3.1+중，측서감정목적기인용양리자지질체전염적방법전염중국창서란소CHO세포。결과PCR확증적특이성편단장도위1 304bp，이차구건중조질립pCDNA3.1+-Enol，측서결과여Genbank중적서원Enol기인cDNA서렬일치。전염중국창서란소세포후가검측도Enol단백적표체。결론구건적진핵표체재체pCDNA3.1+-Enol가재진핵세포내정학표체，저위진일보적역묘연구전정료기출。
Objective To construct the eukaryotic expression vector of mouse alpha-enolase enzyme (Eno1)gene. Methods Enol eDNA of mouse kidney cells was amplified with polymerase chain reaction and inserted into the eukaryotic expression vector of pCDNA3. 1 +. The recombinant plasmid was verified by DNA sequencing and used to transfect Chinese hamster ovary(CHO)cells. Results The amplified fragment was 1 304bp in length. The sequence of Eno1 gene was in accordance with that in Genbank. The Enol protein was detected in CHO cells. Conclusion We successfully constructed the pCDNA3.1 +-Enol eukaryotic expression vector which may pave a way for further studies in vaccine experiment.