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.