WU Xin-Xing, ZHAO Wen-Xian, DING Xiao-Hua, SU Ying-Bin and DING Hong-Zhen. The isolating, cloning and sequencing of HPV 16 E7 gene from cervical carcinoma biopsy DNA of Hubei Province[J]. Virologica Sinica, 1996, 11(3).
Citation: WU Xin-Xing, ZHAO Wen-Xian, DING Xiao-Hua, SU Ying-Bin, DING Hong-Zhen. The isolating, cloning and sequencing of HPV 16 E7 gene from cervical carcinoma biopsy DNA of Hubei Province .VIROLOGICA SINICA, 1996, 11(3) : 220.

湖北地区宫颈癌组织中人乳头瘤病毒16型E7基因的分离、克隆和序列分析

  • 采用加端聚合酶链反应技术,从湖北地区一宫颈癌患者癌组织DNA中分离出人乳头瘤病毒16型(HPV16)E7基因,并在pUC18载体中克隆。经限制性核酸内切酶分析和DNA序列分析,确认了含HPV16E7重组克隆质粒,命名pHPV16E7─HB。DNA序列分析表明,HPV16E7─HB基因全长294bp(与报道的标准株基因长度相同),但其核苷酸顺序中有两处发生了C→T突变,即第43位密码子CAA变为TAA,第76位CGT变为TGT;前者使谷氨酰胺密码子变为终止密码,即无义奕变(nonsensemutation)。这种突变发生在294个碱基的DNA扩增产物之中,不像是PCR本身的错配,而很可能是湖北株与标准株之间的结构差异。

The isolating, cloning and sequencing of HPV 16 E7 gene from cervical carcinoma biopsy DNA of Hubei Province

  • The technique of add-on PCR was used for isolating HPV16 E7 gene from cervical carcinomabiopsy DNA in Hubei province.The E7 gene was inserted into vector pUC18.By restriction en-zyme cleavage and nucleotide sequence detection,a new recombinant plasmid was constructed andnamed it pHPV16 E7-HB. The result of nucleotide sequencing showed that the overall length ofHPV16 E7-HB gene is 294 bp,it is the same as the standard strain.But there were two C→Tmutation in HPV16 E7-HB.Anonsense mutation was identifed at codon 43.a Gln codon(CAA)was converted into ochre termination codon (TAA).The mutations occurring in 294 bp ofDNA PCR production seemed to be a structure difference between HB strain and standard strainrather than a mismatch of PCR itself.

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    The isolating, cloning and sequencing of HPV 16 E7 gene from cervical carcinoma biopsy DNA of Hubei Province

    • 1. Hubei Medical University,Wuhan 430071

    Abstract: The technique of add-on PCR was used for isolating HPV16 E7 gene from cervical carcinomabiopsy DNA in Hubei province.The E7 gene was inserted into vector pUC18.By restriction en-zyme cleavage and nucleotide sequence detection,a new recombinant plasmid was constructed andnamed it pHPV16 E7-HB. The result of nucleotide sequencing showed that the overall length ofHPV16 E7-HB gene is 294 bp,it is the same as the standard strain.But there were two C→Tmutation in HPV16 E7-HB.Anonsense mutation was identifed at codon 43.a Gln codon(CAA)was converted into ochre termination codon (TAA).The mutations occurring in 294 bp ofDNA PCR production seemed to be a structure difference between HB strain and standard strainrather than a mismatch of PCR itself.

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