CAJ | 학술논문

目的：构建精氨酸甲基转移酶2( PRMT2)慢病毒表达载体。方法通过PCR扩增PRMT2 cD-NA,将PRMT2 cDNA连接于GV308载体,经测序确认后,将GV308/PRMT2与pHelper 1.0和pHelper 2.0共转染至293T细胞中,收获病毒,通过Real time PCR测定滴度；将PRMT2慢病毒表达载体侵染293T细胞,通过四环素诱导和Western blot检测PRMT2慢病毒表达载体的表达能力。结果在感染PRMT2慢病毒载体293T细胞中能检测到PRMT2-3Flag融合蛋白的表达。结论成功构建PRMT2的慢病毒表达载体。
목적：구건정안산갑기전이매2( PRMT2)만병독표체재체。방법통과PCR확증PRMT2 cD-NA,장PRMT2 cDNA련접우GV308재체,경측서학인후,장GV308/PRMT2여pHelper 1.0화pHelper 2.0공전염지293T세포중,수획병독,통과Real time PCR측정적도；장PRMT2만병독표체재체침염293T세포,통과사배소유도화Western blot검측PRMT2만병독표체재체적표체능력。결과재감염PRMT2만병독재체293T세포중능검측도PRMT2-3Flag융합단백적표체。결론성공구건PRMT2적만병독표체재체。
Objective To obtain Lentivirus with PRMT2 expression. Methods PRMT2 cDNA was obtained from cDNA pool by RT-PCR,and the PRMT2 cDNA was ligated with GV308 vector; the GV308/ PRMT2 plasmid confirmed by sequencing was cotranfected with pHelper 1. 0 and pHelper 2. 0 into 293T cells to obtain PRMT2 Lentivirus; titer of Lentivirus with PRMT2 expression was assessed by Real Time PCR; the expression of PRMT2 was induced by Doxycycline hyclate and detected by Western blot in PRMT2 Lentivirus infected 239T cells. Results PRMT2 Lentivirus could express PRMT2 in 239T cells. Conclusion Lentivirus with the expression of PRMT2 was successfully established.