Construction and detection of the tissue-specific pINV-HPV16 E6/7 vector

Gao,Hui; Huang,Zhengfang; Shi,Chenlong; Li,Houda

doi:10.3892/ol.2014.2736

Construction and detection of the tissue-specific pINV-HPV16 E6/7 vector

Authors:
- Hui Gao
- Zhengfang Huang
- Chenlong Shi
- Houda Li
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Affiliations: Department of Dermatology, Clinical Medical College, Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225000, P.R. China
Published online on: November 25, 2014 https://doi.org/10.3892/ol.2014.2736
Pages: 857-862

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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.

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