CAJ | 학술논문

目的：研究含登革2型病毒43株（D2-43）NS1基因的重组质粒DNA在幼地鼠肾细胞BHK-21的表达。方法：将含信号肽的NS1基因片段插入到pcDNA3.1的KpnⅠ位点和EcoRⅠ位点之间，获得重组表达载体pcDNA-NS1。用电穿孔法将其导入BHK-21细胞，G418选择培养。挑取单细胞克隆，RT-PCR及蛋白质印迹法鉴定NS1基因的稳定表达。结果：在随机挑取的5个单细胞克隆中，有4个克隆的RT-PCR鉴定为阳性，蛋白质印迹结果表明NS1基因获得表达。结论：构建的pcDNA-NS1质粒在BHK-21细胞中有稳定表达，因此含NS1基因的该重组质粒DNA可用作核酸免疫。
목적：연구함등혁2형병독43주（D2-43）NS1기인적중조질립DNA재유지서신세포BHK-21적표체。방법：장함신호태적NS1기인편단삽입도pcDNA3.1적KpnⅠ위점화EcoRⅠ위점지간，획득중조표체재체pcDNA-NS1。용전천공법장기도입BHK-21세포，G418선택배양。도취단세포극륭，RT-PCR급단백질인적법감정NS1기인적은정표체。결과：재수궤도취적5개단세포극륭중，유4개극륭적RT-PCR감정위양성，단백질인적결과표명NS1기인획득표체。결론：구건적pcDNA-NS1질립재BHK-21세포중유은정표체，인차함NS1기인적해중조질립DNA가용작핵산면역。
Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.