عنوان به زبان ديگر :
Cloning and Expression of Hepatitis C Virus Core Protein in pGemex-1 Expression Vector
پديد آورندگان :
Kazemi Bahram نويسنده Kavakeb P , Bandehpour Mojgan نويسنده , Seyed Negar نويسنده , Roozbehi Mehrnaz نويسنده , Mosaffa Nariman نويسنده
چكيده لاتين :
Background: Hepatitis C virus is one of the main causes of chronic hepatitis in developing
countries. There are 170 million affected people around the world as reported by the World Health
Organization. The treatment of hepatitis C is not successful in most cases; it is extremely costly,
and requires prolonged therapy, therefore it is desirable to develop a vaccine to prevent the spread
of hepatitis C virus.
Methods: Hepatitis C virus RNA was extracted from a hepatitis C virus-infected serum sample.
cDNA was synthesized and the hepatitis C virus core gene was amplified by polymerase chain
reaction. The polymerase chain reaction product was cloned in pGEMEX-1 expression vector and
expressed in Escherichia coli BL21 strain with DE3 (a ë prophage carrying the T7 RNA
polymerase gene) host by induction of promoter using one millimolar isopropyl â-Dthiogalactopyranoside
in laboratory scale. Induced lysate cells were electrophoresed on SDS-
polyacrylamide gel.
Results: A protein band was detected in induced cells in comparison with non-induced cells.
Expressed protein was confirmed by gel diffusion and dot blot analysis using induced lysate cells
as antigen and hepatitis C virus-infected serum as antibody.
Conclusion: In present study, we have provided a recombinant plasmid based on hepatitis C
virus core gene.