CAJ | 학술논문

目的：通过不同方案包装HIV-1假病毒，确定获得高重复性、高稳定性和高滴度假病毒的方法。方法从转染效率影响因素出发，通过不同的转染方案，利用pNL4-3 LucR-E-质粒与VSV-G质粒共转染293细胞包装病毒，分别于48 h和72 h收取假病毒并重新感染293细胞，48 h后测荧光素酶报告基因。结果成功包装出HIV-1假病毒，不同包装方案的病毒滴度不同，测得报告基因最高接近107 RLUs。结论应用PEI转染试剂、大提质粒pNL4-3︰VSV-G为4︰1时，转染效率最高，包装的假病毒滴度最高。
목적：통과불동방안포장HIV-1가병독，학정획득고중복성、고은정성화고적도가병독적방법。방법종전염효솔영향인소출발，통과불동적전염방안，이용pNL4-3 LucR-E-질립여VSV-G질립공전염293세포포장병독，분별우48 h화72 h수취가병독병중신감염293세포，48 h후측형광소매보고기인。결과성공포장출HIV-1가병독，불동포장방안적병독적도불동，측득보고기인최고접근107 RLUs。결론응용PEI전염시제、대제질립pNL4-3︰VSV-G위4︰1시，전염효솔최고，포장적가병독적도최고。
Objective To package HIV-1 pseudotyped virus through different ways, to define methodology of packaging high reproductive, high stable and high titer pseudotyped virus. Methods In view of the transfection efifciency factors, through different transfection ways, pNL4-3 LucR -E -plasmid and VSV-G plasmid were transfected into 293 cells packaging virus, and the HIV-1 pseudotyped virus after 48 h and 72 h were collected, respectively, then reporter gene after infecting 293 cells for 48 h were detected. Results HIV-1 pseudotyped virus were packaged successfully, the virus titer differs from packaging ways, and reporter gene measured up to 107 RLUs. Conclusion When using PEI transfection reagent, and plasmid pNL4-3︰VSV-G as 4︰1, the highest transfection efifciency and the highest viral titer could received.