钟警,杨心治,张艳敏,高海花,秦旭平,文格波.PRMT2慢病毒表达载体的构建及鉴定.[J].中南医学科学杂志.,2015,43(1):73-77.
PRMT2慢病毒表达载体的构建及鉴定
The Establishment and Identification of Lentivirus with PRMT2 Expression
投稿时间:2014-09-04  
DOI:
中文关键词:  PRMT2  慢病毒载体  基因治疗
英文关键词:PRMT2 lentivirus gene therapy
基金项目:国家自然科学基金青年项目(31200573)、国家自然科学基金面上项目(81272906)和湖南省自然科学基金(12JJ3116)
作者单位
钟警,杨心治,张艳敏,高海花,秦旭平,文格波* (南华大学附属第一医院临床医学研究所湖南 衡阳 421001) 
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中文摘要:
      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PRMT2cDNA,将PRMT2cDNA连接于GV308载体,经测序确认后,将GV308/PRMT2与pHelper1.0和pHelper2.0共转染至293T细胞中,收获病毒,通过RealtimePCR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Westernblot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。
英文摘要:
      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.
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