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HPV6晚期表达基因L1的克隆并在杆状病毒表达系统的表达及其抗血清制备

Cloning,Expression of HPV6 L1 in Bac-to-Bac Baculovlovirus Expression System and Preparation of anti-r-HPV6 L1 Serum
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摘要 目的 获得具有正确序列的人乳头瘤病毒 HPV6型晚期表达基因 L1 ,并获得其高活性表达 L1基因特异性抗血清 ,为进一步研究及临床检验应用奠定物质基础。方法 用 PCR法从感染人乳头瘤病毒患者尖锐湿疣组织中获得 HPV6晚期表达基因 L1 ,测序验证后 ,将正确的 HPV6晚期表达基因 L1序列 ,插入 Bac- to- Bac杆状病毒表达系统 ,转染昆虫细胞 Sf9,表达外壳蛋白 L 1 ,并以之作为抗原免疫兔 ,获得特异性抗血清。结果 获得了正确序列 HPV6晚期表达基因 L1 ,在 Bac- to- Bac杆状病毒系统活性表达并获得其特异性抗血清。结论 获得 HPV6晚期表达基因 L1 ,及有抗原活性的表达和相应抗血清。 Objective Human Papillomavirus 6 L1 gene was cloned from condyloma acuminata and expressed actively in Sf9 insect cells by Bac-to-Bac Baculovirus expression vector system.Methods Human Papillomavirus6 L1 gene was acquired by PCR from CA.At first,Human Papillomavirus type6 L1 gene was insert into pFast Bac vector,then the recombiant pFast Bac-Angio(K1-4) transfect competent DH10 Bac cells to construct recombiant HPV6 L1 Bacmid DNA,Sf9 cells were cotrasfected with recombiant HPV6 L1-Bacmid DNA using cellfectin.Then recombiant HPV6 L1 was used as antigen to immunity rabbit.Results After 4 round amplification of recombiant baculovirus,high expression level of HPV6 L1 in Sf9 cells infected was detected by immunoblotting with HPV6 L1 antibody,and anti-reHPV6 L1 was acquired.The usitility in diagniosis and therapy need further study.Conclution HPV6 L1 genewas acquired and actively expressed in Bac-to-Bac Baculovirus expression system.The anti-HPV6 L1 serum has been acquired.
出处 《陕西医学检验》 2002年第1期1-3,共3页 Shaanxi Journal of Medical Laboratory Sciences
关键词 HPV6晚期表达基因L1 克隆 Bac-to-Bac杆状病毒表达系统 抗血清 human papillomavirus cloning Bac-to-Bac Baculovirus expression system
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  • 1zur Hausen H. Papillomaviruses in human cancers[J].Proc Assoc Am Physicians,1999;111(6):581~587.
  • 2Nagai Y,Maehama T,Asato T,et al.Detection of Human Papillomavirus DNA in Primary and Metastatic Lesions of Carcinoma of the Cervix in Women from Okinawa,Japan [J].Am J Clin Oncol, 2001;24(2):160~166.
  • 3Caparros-Wanderley W,Savage N,Hill-Perkins M,et al.Intratype sequence variation among clinical isolates of the human papillomavirus type 6 L1 ORF:clustering of mutations and identification of a frequent amino acid sequence variant [J].J Gen Virol,1999;80(Pt 4):1025~1033.
  • 4Schwarz E,Durst M,Demankowski C,et al.DNA sequence and genome organization of genital human papillomavirus type 6b[J].EMBO J,1983;2(12):2341~2348.
  • 5Luckow VA,Stephen CL,Gerard FB,et al.Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes in to a baculovirus genome propagated in E.Coli[J].J Virology,1993;67(8):4566~4579.
  • 6Hartig PC,Cardon MC.Rapid efficient production of baculoviru expression vectors[J].J Viro Method,1992;38(1):61~70.
  • 7Lo KW,Cheung TH,Chung TK,et al.Clinical and prognostic significance of human papillomavirus in a chinese population of cervical cancers[J].Gynecol Obstet Invest,2001;51(3):202~207.
  • 8Hove MG,Hightower BJ,Graves K.Use of combining in situ hybridization and immunohistochemistry in detecting human papillomavirus on routine sections in cases of diagnostic uncertainty[J].Cent Afr J Med,2000;46(8):217~221.
  • 9Banks L,Matlashewski G,Pim D,Churcher M,et al.Expression of human papillomavirus type 6 and type 16 capsid proteins in bacteria and their antigenic characterization[J].J Gen Virol,1987;68:3081~3089.
  • 10Robert C,Rose R C,Reichman W.Human papillomavirus type Ⅱrecombinant virus-like particles induce the formation of neutralizing antibodies and detect HPV-specific antibodies in human sera[J].J Gen Virol,1994;75:2075~2079.
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