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肝细胞癌组织中HBx基因缺失型突变体真核表达载体的构建和鉴定 被引量：6

Construction of Eukaryotic Expression Recombinant Plasmids for the Expression of Delete Mutated HBx
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摘要 目的构建缺失型突变体HBx-d382和HBx-d431真核表达载体。方法以pGM-T/HBx-d382和pGM-T/HBx-d431质粒为模板,PCR扩增含KpnⅠ和ApaⅠ酶切位点的HBx基因片段。双酶切后再与载体pcDNA3载体重组连接。重组质粒转化到大肠杆菌DH5α后,经酶切鉴定和DNA序列测定,筛选出重组pcDNA3/HBx-d382和pcDNA3/HBx-d431真核表达载体。结果成功构建了重组pcDNA3/HBx-d382和pcDNA3/HBx-d431真核表达载体,经DNA序列测定与比对,重组前后HBx基因片段序列一致。结论pcDNA3/HBx-d382和pcDNA3/HBx-d431真核表达载体构建成功,可为HBx基因缺失型突变体,特别是HBx-d382基因的生物学功能研究提供基因材料。 Objective To construct the eukaryotic expression recombinant plasmid containing HBx-d382 and HBx-d431 genes, and explore the relationship between HBx deletion mutant and hepatocellular carcinoma. Method HBx-d382 and HBx-d431 genes were PCR amplified, using specific primers containing Kpn I and Apa I restriction sites, from pGM-T/HBx-d382 and pGM-T/HBx-d431 plasmids DNA, and then subcloned into pcDNA3 to generate pcDNA3/HBx-d382 and pcDNA3/HBx-d431 recombinant plasmids. Recombinant plasmids were identified by Kpn I and Apa I restriction analysis and DNA sequencing. Result We have successfully constructed the eukaryotic expression vector for the expression of HBx-d382 and HBx-d431. Conclusion pcDNA3/HBx-d382 and pcDNA3/ HBx-d431 eukaryotic expression recombinant plasmids can be used for functional studies of HBx-d382 and HBx-d431 genes in carcinogenesis of hepatocellular carcinoma.
作者 朱平安 谭德明 陈莉 彭忠田 宋琳
机构地区 深圳市第七人民医院 中南大学湘雅医院传染科
出处 《热带医学杂志》 CAS 2008年第4期332-334,361,共4页 Journal of Tropical Medicine
关键词 HBX基因 肝细胞癌 乙型肝炎病毒 缺失型突变体 真核表达载体 HBx gene hepatocellular carcinoma hepatitis B virus deletion mutant eukaryotic expression vector
分类号 R735.7 [医药卫生—肿瘤]
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参考文献17

  • 1Moradpour D, Blum HE. Pathogenesis of hepatocellular carcinoma [J]. Eur J Gastroenterol Hepatol, 2005,17 (5) : 477-483.
  • 2Chung TW, Lee YC, Ko JH, et al. Hepatitis B virus X protein modulates the expression of PTEN by inhibiting the function of p53, a transcriptional activator in liver cells [J]. Cancer Research, 2003,63 ( 13 ) : 3453-3458.
  • 3Kwun H J, Jang KL. Natural variants of hepatitis B virus X protein have differential effects on the expression of cyclin- dependent kinase inhibitor p21 gene [J]. Nucleic Acids Research, 2004,32(7) :2202-2213.
  • 4Ogden SK, Lee KC, Barton MC. Hepatitis B viral transactivator HBx alleviates p53-mediate drepression of alpha-fetoprotein gene expression [J]. J Biol Chem, 2000,275 (36) :27806- 27814.
  • 5Su F,Theodosis CN ,Schneider RJ. Role of NF-kappaB and myc proteins in apoptosis induced by hepatitis B virus HBx protein [J]. J Virol, 2001,75(1 ):215-225.
  • 6Shama YM, Barak O, Doitsh G, et al. Hepatitis B virus pX interacts with HBXAP,a PHD finger protein to coactivate transcription [J ]. J Biol Chem, 2002,277 (12) : 9982-9988.
  • 7lavarone M, Trabut JB, Delpuech O, et al. Characterisation of hepatitis B virus X protein mutants in tumour and non-tumour liver cells using laser capture microdissection [J ]. J Hepatol, 2003,39(2) :253-261.
  • 8Tu H, Bonura C, Giannini C, et al. Biological impact of natural COOH-terminal deletions of hepatitis B virus X protein in hepatocellular carcinoma tissues [J]. Cancer Res, 2001,61 (21) :7803-7810.
  • 9Ping'an ZHU,Deming TAN,Zhongtian PENG,Fei LIU,Lin SONG.Polymorphism Analyses of Hepatitis B Virus X Gene in Hepatocellular Carcinoma Patients from Southern China[J].Acta Biochimica et Biophysica Sinica,2007,39(4):265-272. 被引量：29
  • 10Ahn JY, Jung EY, Kwun HJ, et al. Dual effects of hepatitis B virus X protein on the regulation of cell-cycle control depending on the status of cellular p53 [J]. J Gen Virol, 2002,83 (Ptl 1 ) :2765-2772.

二级参考文献41

  • 1林静,朱明华,曲建慧,李芳梅,倪灿荣.乙型肝炎病毒X基因经p53依赖性与非依赖性途径对p21^(WAF1)表达的影响[J].癌症,2004,23(7):749-755. 被引量：9
  • 2杨少波,王孟薇,尤纬缔,于国.人肝癌c─myc基因产物和HBxAg表达研究[J].军医进修学院学报,1996,17(2):110-112. 被引量：1
  • 3Birrer RB, Birrer D, Klavins JV. Hepatocellular carcinoma and hepatitis virus [J]. Ann Clin Lab Sci, 2003,33( 1 ):39-54.
  • 4Tu H, Bonura C, Giannini C, et al. Biological impact of natural COOH-terminal deletions of hepatitis B virus X protein in hepatocellular carcinoma tissues [J]. Cancer Res, 2001,61(21) :7803-7810.
  • 5Sirma H, Giannini C, Poussin K, et al. Hepatitis B virus X mutants, present in hepatocellular carcinoma tissue abrogate both the antiproliferative and transactivation effects of HBx [J].Oncogene, 1999,18(34) :4848-4859.
  • 6Iavarone M, Trabut JB, Delpuech O, et al. Characterisation of hepatitis B virus X protein mutants in tumour and non-tumour liver cells using laser capture microdissection [J]. J Hepatol,2003,39(2): 253-261.
  • 7Murakami S, Cheong JH, Kaneko S. Human hepatitis virus X gene encodes a regulatory domain that represses transactivation of X protein [J]. J Biol Chem, 1994,269(21): 15118-15123.
  • 8Elmore LW, Hancock AR, Chang SF, et al. Hepatitis B virus X protein and p53 tumor suppressor interactions in the modulation of apoptosis [J]. Proc Natl Acad Sci USA, 1997,94(26): 14707-14712.
  • 9Feitelson MA, Clayton MM. X antigen/antibody markers in hepadnavirus infections. Antibodies to the X gene product (s)[J]. Gastroenterology, 1990,99(2): 500-507.
  • 10Shirakata Y, Kawada M, Fujiki Y, et al. The X gene of hepatitis B virus induced growth stimulation and tumorigenic transformation of mouse NIH3T3 cells [J]. Jpn J Cancer Res,1989,80(7): 617-621.

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热带医学杂志
2008年 第4期

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