[1]李朵璐),余祖江),阚全程)#.反义乙型肝炎病毒S基因T载体克隆的构建*[J].郑州大学学报(医学版),2009,(05):963-965.
 LI Duolu),YU Zujiang),KAN Quancheng).Construction and identification of antisense HBV S gene cloning by T vector[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(05):963-965.
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反义乙型肝炎病毒S基因T载体克隆的构建* ()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年05期
页码:
963-965
栏目:
论著
出版日期:
2009-09-01

文章信息/Info

Title:
Construction and identification of antisense HBV S gene cloning by T vector
作者:
李朵璐1)余祖江2)阚全程1)#
1)郑州大学第一附属医院药剂科郑州4500522)郑州大学第一附属医院感染科郑州450052
Author(s):
LI Duolu1)YU Zujiang2)KAN Quancheng1)
1)Department of Pharmacy,the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 2)Department of Infectious Disease,the First Affiliated Hospital,Zhengzhou University, Zhengzhou 450052
关键词:
肝炎病毒乙型S 基因反义T载体
Keywords:
hepatitis B virusS geneantisenseT vector
分类号:
R373.2
摘要:
构建含部分乙型肝炎病毒(HBV)S基因反义序列的基因克隆。方法:根据GenBank中HBVS基因序列和计算机软件mCLONE分析的结果,设计S基因2侧引物,直接扩增出适合长度的反义HBVS基因,与PMD18T载体连接后转化DH5α,筛选阳性克隆,抽提重组质粒并进行PCR及酶切鉴定,再行序列分析。结果:获得226bp含限制性内切酶位点的阳性产物,T载体克隆经PCR及酶切鉴定和序列分析后证实,克隆片段的反向序列与GenBank中该基因的序列同源性为97%。结论:成功构建了含反义HBVS基因部分序列的T载体克隆。
Abstract:
To construct the PMD18T vector containing part of the antisense HBV S gene.Methods:Sense and antisense primers of HBV S gene were designed according to the sequence of the HBV S gene from GenBank and the analytical results of the mCLONE software. Using the primers, the proper length antisense HBV S gene was obtained by PCR. The amplified DNA fragments were ligated into PMD18T vector, and then transformed into DH5α.The positive clones were screened out and the recombinant plasmids were isolated from such clones. The recombinants were identified by PCR, restriction endonucleases analysis and sequencing.Results: PCR assay showed that a DNA fragment of 226 bp was amplified from the recombinant plasmids. Restriction endonucleases digestion confirmed the target fragment had been inserted into PMD18T successfully. The sequence results verified that the antisense sequence of the target fragment had high homology (97%) with the HBV S gene sequence in GenBank.Conclusion: The T vector clone with part of the antisense HBV S gene has been constructed successfully.

参考文献/References:

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[2]Chu CM, Liaw YF.Natural history of chronic hepatitis B virus infection: an immunopaththological study[J]. J Gastroenterol Hepatol,1997,13(9/10):5 218
[3]Chu CM.Natural history of chronic hepatitis B virus infection in adults with emphasis on the occurrence of cirrhosis and hepatocellular carcinoma[J]. J Gastroenterol Hepatol,2000,15(suppl):E25
[4]Tan A,Yeh SH,Liu CJ,et al.Viral hepatocarcinogenesis: from infection to cancer[J]. Liver Int,2008,28(2):175
[5]GonzalezAsequinolaza G,Crettaz J,Ochoa L,et al.Gene therapy for viral hepatitis[J]. Expert Opin Biol Ther,2006,6(12):1 263
[6]Xu R,Cai K,Zheng D,et al.Molecular therapeutics of HBV[J].Curr Gene Ther,2003,3(4):341

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30472031 #通讯作者,男,1963年生,博士,教授,研究方向:感染性疾病的基因治疗与免疫药理,Email:kanqc@yahoo.com.cn
更新日期/Last Update: 2010-05-14