12. PCR AMPLIFICATION AND EXPRESSION ANALYSIS OF P7 GENE OF HCV IN Huh-7 CELL LINE

Shazia Tariq1, Tahir Sarwar2, Jawad Ahmed2, Ghazala Afridi2, Shenaz Bukhtiar3, Sunia Arif3, Qurat-ul-Ain Tariq3, Hayyan3

1Department of Pathology, Gajju Khan Medical College, Swabi - Pakistan
2Department of Pathology, Institute of Basic Medical Sciences, Khyber Medical University, Peshawar - Pakistan
3Department of Pathology, Hayatabad Medical Complex, Peshawar - Pakistan

Abstract

Objectives: To know the PCR amplification and expression analysis of p7 gene of HCV in Huh-7 cell line.
Material & Methods: In this study, blood samples were collected from HCV positive patients from Peshawar Khyber
Pakhtunkhwa, Pakistan. RNA was extracted and then converted into cDNA using gene specific primer. P7 gene of HCV
was amplified using PCR and subsequently sequenced through Sanger sequencing. P7 gene was then cloned into
mammalian expression vector pCR3.1and expressed in Huh-7 cell line. RNA was extracted from transfected cells and
then converted into cDNA with the oligo (dt) primers, expression of p7 gene was studied.
Results: For the first ever time in KP, Pakistan, p7 gene was amplified, sequenced, cloned and expressed in Huh 7 cell line.
Conclusion: Expression of p7 gene of HCV can be used for comparative analysis of another strain of HCV of different
location and this provides an opportunity for the development of novel anti-HCV drugs with good efficacy and minimal
side-effects.

Keywords: RT (reverse transcriptase), Hepatitis c virus, Untranslated regions, human hepatocellular7- cell line

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