Cloning hepatitis C virus E2 gene full-length from genotype 2a (JFH1) to pET28a (+) vector and transformation of an Escherichia coli DH5α strain

Aim and The envelope glycoprotein E2 in Hepatitis C virus is required for host-cell entry. The nucleotide sequence diversity in carboxyl-terminus (C-terminal) of this gene results in induction of neutralizing antibody that makes this gene an important target for vaccine development studies. In this study we tried to make a recombinant pET28a () containing full length of the E2 gene from genotype 2a (JFH1) followed by being transformed into the E-coli (DH5a).
Material and Full-length of HCV E2 region from genotype 2a (JFH1) was amplified by PCR reaction using the specific primers including the restriction sites for EcoR1 and Nde1. The confirmed PCR product was cloned into the pET28a () vector and transformed into the E-coli (DH5a) using the heat shock method.
The HCV E2 gene was successfully cloned into the pET28a () vector in full length size. Transformation of the E-coli strain DH5a by this recombinant vector was confirmed.
Transformation of the E-coli (DH5a) using the recombinant vector pET28a- HCV 2a (JFH1) E2 gene provides useful tools for further expression of this gene in full length size in different expression system.