CAJ | 학술논문

目的：研究慢病毒在感染细胞后，是否全部整合入染色体。方法用慢病毒载体的假病毒感染293 T细胞后，抽提培养2 d和10 d的细胞基因组DNA。通过定量PCR检测细胞基因组中LTR拷贝数，计算出整合率。分别取病毒感染3 d和11 d的细胞，离心重悬后取部分细胞放在荧光显微镜下拍照，在蛋白水平上验证此结论。结果①慢病毒载体在宿主细胞基因组内的平均整合率（47±16）％（n＝10）；②细胞感染慢病毒后3 d和11 d的GFP照片也证实了慢病毒在宿主细胞基因组内部分整合。结论慢病毒载体的假病毒感染293 T细胞后，仅有部分的载体骨架是整合入基因组中的，整合入293 T细胞基因组中的外源基因能够较长时间稳定表达。
목적：연구만병독재감염세포후，시부전부정합입염색체。방법용만병독재체적가병독감염293 T세포후，추제배양2 d화10 d적세포기인조DNA。통과정량PCR검측세포기인조중LTR고패수，계산출정합솔。분별취병독감염3 d화11 d적세포，리심중현후취부분세포방재형광현미경하박조，재단백수평상험증차결론。결과①만병독재체재숙주세포기인조내적평균정합솔（47±16）％（n＝10）；②세포감염만병독후3 d화11 d적GFP조편야증실료만병독재숙주세포기인조내부분정합。결론만병독재체적가병독감염293 T세포후，부유부분적재체골가시정합입기인조중적，정합입293 T세포기인조중적외원기인능구교장시간은정표체。
Objective To study how much infected Lentiviral Vectors are integrated into the host cell genome?Methods The lentiviral pro?viral copy numbers in the infected 293 T cells chro?mosome was determined by quantitative PCR analysis?The ratio of the integration on day 10 over day 2 infection was used to determine the integration rate?Moreover, day 11 and day 3 infection pictures were used to prove the conclusion?Results The integration rate on average was :47% ± 0?16 ( n=10 );The GFP pictures taken on day 3 and day 11 proved such a conclusion?Conclusion Only part of infected Lentiviral Vectors are integrated into the host cell genome, and the integrated trans?gene could relatively stable express its transgene.