以HIV-1为骨架的慢病毒载体在其假病毒的转录本中保留其内含子不被剪切  

HIV-1 Based Lentiviral Vector Protects its Transgene Introns from Splicing in its Pseudovirus

作　　者：张帆[1,2] 方彧聃[1,2] 张敬之[1,2] 

机构地区：[1]上海交通大学医学遗传研究所 [2]上海交通大学附属儿童医院,中国上海200040

出　　处：《生命科学研究》2011年第3期195-202,共8页Life Science Research

基　　金：国家自然科学基金资助项目(30870943);上海市自然科学基金资助项目(11ZR1429900)

摘　　要：通过在FUGW中克隆入各外源基因表达盒,如红系特异表达人β珠蛋白(HBG),或山羊乳腺特异表达人血清白蛋白(pBLG-ALB)等,研究以HIV-1为骨架的慢病毒载体FUGW在其假病毒的转录本中内含子的剪切情况.将慢病毒载体制备成假病毒后抽提其病毒RNA,通过逆转录和PCR验证其内含子剪切保留情况;同时,在相对应的慢病毒假病毒介导的转基因小鼠中也进行类似的PCR验证.在制备假病毒的转录过程中,其外源基因所携带的內含子在病毒所包含的基因组RNA中被部分保护不被剪切,且优先保护5′端的内含子.此现象正好与野生型HIV-1的保护顺序相反.To study the fate of transgene introns in the pseudovirus of HIV-1 based lentiviral vector,FUGW,human β-globin promoter and gene（HBG）,or human serum albumin（pBLG-ALB） gene driven by goat β-lactoglobulin promoter were cloned into FUGW,and their pseudoviruses were then prepared.The pseudovirus RNA was undergone reverse transcription followed with PCR amplification in order to study their introns.Simultaneously,genomic DNA of lentiviral vector mediated transgenic mice was also used for PCR amplification to determine the transgene introns.It was found that the transgene introns were protected from splicing in both pseudoviruses and transgenic mice.The splicing order was 3′intron prior to 5′intron in the lentiviral pseudoviruses,which was in contrast to the case reported in HIV-1.

关 键 词：艾滋病毒 慢病毒载体 内含子 剪切 剪切保护 

分 类 号：Q752[生物学—分子生物学]