Expression of Bovine leukemia virus p24 Protein in Bacterial Cell

Abstract

Bovine leukemia virus (BLV) is a member of the family Retroviridae, genus Deltaretrovirus that has three important gene including gag, pol and env. This virus causes B-cell lymphocytosis and lymphosarcoma in cows. In the first step PCR product of gag gene of BLV isolated in different regions of Iran and BLV-FLK strain were cloned in to a pTZ57R/T vector, then insert were digested by BglII and XhoI restriction enzymes and cloned in to pET-28(a) as an expression vector. For the expression of p24 protein, the pET-28(a) recombinant vector was transformed in BL21(DE3) strain of E. coli competent cell and after induction of the protein having been expressed by IPTG, the presence of gag expressed protein was shown in immunoblotting and SDS-PAGE system. With respect to the remarkable frequency of infection to BLV in Iran and the necessity of controlling it through vaccination with recombinant vaccines of gp51, manufacturing and applying the recombinant p24 protein are vital goals in recognition and distinction between infection and responses caused by vaccine.

Molecular detection of bovine leukemia virus in peripheral blood of Iranian cattle, camel and sheep

“…The serological tests used for diagnosis of bovine leukemia include the Agar Gel Immuno Diffusion (AGID) and the Enzyme-Linked Immuno Sorbent Assay (ELISA) (Momtaz et al 2008, OIE 2012. Both tests are less sensitive when compared to polymerase chain reaction (PCR) described by several researchers (OIE 2012).…”

Expression of p24 gag Protein of Bovine Leukemia Virus in Insect Cells and Its Use in Immunodetection of the Disease

“…The expression of proteins from different viruses, such as bovine immunodeficiency virus [23] and bovine respiratory syncytial virus [24], has been already described. In the last decade, BLV gag proteins p24 have been expressed in heterologous expression systems such as Escherichia coli [25][26][27][28] or Saccharomyces cerevisiae [29]. In a direct comparison study for recombinant protein production in the E. coli, Pichia pastoris, Saccharomyces cerevisiae and Spodoptera frugiperda systems, Christopher et al [30] reported that baculovirus-mediated expression in Sf21 cells is the most efficient system capable of producing large amounts of recombinant protein.…”