Evaluation of Phycocyanin Promoter Function in Bacteria by Investigating the Expression of HBsAg

Document Type : Original Article

Authors

1 1. Razi Herbal Medicines Research Center, Lorestan University of Medical Science, Khorramabad, Iran 2. Biotechnology Incubator Center, North Research Center, Pasteur Institute of Iran, Amol, Iran

2 3. Department of Plant Breeding & Biotechnology. SANRU, Sari, Iran

3 4. Agricultural & Natural Resources College, University of Tehran, Iran

4 1. Razi Herbal Medicines Research Center, Lorestan University of Medical Science, Khorramabad, Iran

5 5. Department of Cellular and Molecular Biology, Babol Branch, Islamic Azad University, Babol, Iran

6 7. Department of Plant production and genetic engineering. Faculty of agriculture. Lorestan University, Iran.

Abstract

Many items affect the yields of every recombinant protein production. Promoters are one of the key regulatory elements which control the level of recombinant protein expression in the host. Although in some studies (Hepatitis B Surface Antigen) HBsAg was cloned in E. coli, in many of them common promoters were used. In this study two co-vectors (PHK and PHGK) were designed and used. Both of them were shuttle types based on phycocyanin-specific promoter and each was a combination of two types of vectors: a plasmid and a transposon.  ELISA results, in line with Western blot results, showed the ability of phycocyanin promoter as well as revealed the expression level of HBsAg in transgenes from the Top10 PHK vector is higher than those of Top10 PHGK. The expression of HBsAg under the phycocyanin promoter in the present study is 7.5 µg/lit. In the current study as a pilot step, our attitude and objective of bacteria expression are to evaluate the function of the phycocyanin promoter based on the HBsAg gene. Due to the nature of the shuttle vector, in continuation of the present study to reduce the non-responsive populations, high antigen expression, meet the need for renewal and, other benefits, expression of HBsAg in other than bacterial host is under investigation.

Keywords

Main Subjects