乙型肝炎病毒前-S2蛋白反式激活蛋白1基因的克隆化研究  

Cloning of human gene 1 transactivated by pre-S2 protein of hepatitis B virus

作　　者：纪冬[1,2] 刘妍[2] 韩萍[1] 陈国凤[1] 辛绍杰[2] 

机构地区：[1]解放军军医进修学院,北京100853 [2]解放军第302医院,北京100039

出　　处：《军医进修学院学报》2010年第1期55-57,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：国家自然科学基金(30700713);军队"十一五"课题(06MA361)~~

摘　　要：目的应用基因表达谱芯片技术及生物信息学技术筛选并克隆乙型肝炎病毒(HBV)前-S2蛋白反式激活新型靶基因,进一步阐明HBV感染相关性疾病的发病机制。方法以HBV前-S2蛋白表达质粒pcDNA3.1(-)-PreS2转染HepG2细胞,以空载体pcDNA3.1(-)为平行对照,提取总RNA进行基因表达谱芯片分析。并克隆HBV前-S2反式激活作用的新的靶基因。结果获得该新基因的全长序列,并测序证实,因其可以被前-S2蛋白反式激活,故命名为前S2反式激活蛋白1(PS2TP1),已在GenBank中注册,注册号:AY561706。PS2TP1基因的编码序列全长为1113个核苷酸(nt),编码产物由370个氨基酸残基(aa)组成。结论HBV前-S2蛋白具有反式激活功能,调节宿主细胞某些基因的表达,从而改变宿主正常的应答水平,引起病变。Objective To screen and clone the target genes transactivated by hepatitis B virus （HBV） Pre-S2 protein in order to pave a way for elucidating the pathogenesis of HBV infection. Methods HBV Pre-S2 coding DNA fragment was amplified by polymerase chain reaction （PCR） using G3767 plasmid containing the full length of HBV genome as a template. An expressive vector of pcDNA 3.1 Pre-S2 was constructed with routine molecular biological methods. HepG2 cells were transfected with pcDNA3.1 （-） and pcDNA3.1 Pre- S2, respectively, using the FuGENE6 transfeetion reagent. Total RNA was isolated and reverse transcribed, eDNAs were subjected to mieroarray screening with 1 152 cDNA probes. Results Full length sequences of the obtained genes were searched from GeneBank. One of the obtained genes was a new gene with unknown functions. The new gene that has no homology with known genes in GeneBank was confirmed with its full length DNA cloned by polymerase chain reaction （PCR）. Reverse transcription PCR （RT-PCR） was used to amplify the new gene, PS2TP1, from mRNA of HepG2 cells （The access number of PS2TP1 gene Gen-Bank is AY561706）. Conclusion HBV Pre-S2 protein is a transcriptional transaetivator. The results of our study may help to find the molecular mechanism underlying the transaetivating effects of HBV Pre-S2 protein and to develop new therapies for chronic hepatitis B.

关 键 词：嗜肝DNA病毒属 蛋白 反式激活遗传学 基因 克隆 分子 

分 类 号：R512.62[医药卫生—内科学]