Abstract
A tissue-specific promoter can control downstream gene expression in tissues or organs. The human involucrin (hINV) promoter (pINV) that contains 2474 bp of hINV upstream sequence is able to regulate tissue-specific gene expression. This tissue specificity may be important for the prevention and treatment of human papilloma virus infections. pINV was cloned by polymerase chain reaction and the human papillomavirus (HPV)16 E6/7 gene was obtained from the cancer tissue samples of patients with cervical carcinoma at the Yangzhou Maternal and China Health-Care Center of Jinagsu Province (Yangzhou, China). First, specific primers were designed according to the genomic DNA sequence of the HPV16-type standard strain that has been reported and the E6/7 gene was acquired by PCR. The carcinogenic fraction of the E6/7 gene was removed and the remaining section was cloned into T vectors, sequenced correctly and then cloned into the eukaryotic expression vector pCEP4, which was lacking the CMV promoter. The positive recombinants were identified using blue-white screening and endonuclease digestion, subsequent to sequencing and analysis, and the tissue-specific recombinant pINV-HPV16E6/7 plasmids was detected.
Highlights
Cervical cancer is the second most common type of cancer in females worldwide, in numerous developing countries [1]
The carcinogenic fraction was removed from the E6/7 gene and the remaining section was cloned into T vectors, correctly sequenced and cloned into the eukaryotic expression vector pCEP4, which was separated from the CMV promoter
The tissue‐specificity of the pINV was judged through the detection of HPV16 E6/7 expression in various types of cells
Summary
Cervical cancer is the second most common type of cancer in females worldwide, in numerous developing countries [1]. To reduce the high morbidity rate, the development of effective prevention and gene therapy is needed. With the advancement of molecular biology, gene therapy can be developed for a variety of cancers [1,2]. Human papillomaviruses (HPVs) are double‐stranded DNA viruses and, at present, >200 genotypes have been identified [3,4]. The tissue specificity of the recombinant pINV‐HPV16E6/7 plasmid was detected in the present study
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