the objective of this research was to investigate the influence of the over-expression of recombinant interferon-g during high cell density cultivation on cellular characteristics of recombinant e. coli. batch and fed-batch culture techniques were employed to grow escherichia coli bl21 for production of human gamma-interferon in pet expression system. final cell densities in batch and fed-batch...

background: the aim of this study was to produce a stable cho cell line expressing tenecteplase. materials and methods: in the first step, the tenecteplase coding sequence was cloned in a pdb2 vector containing attb recognition sites for the phage φc31 integrase. then, using lipofection, the cho cells were co-transfected with constructed recombinant plasmid encoding tenecteplase and attb recogn...

rotavirus nonstructural glycoprotein nsp4 can induce diarrhea in newborn mice. it has been suggested that nsp4 may be a key determinant for rotavirus pathogenesis and a target for vaccine development. in order to study the biological and morphological role of nsp4 a large amount of the purified protein and antibody against it are required. simian rotavirus sa11 was propagated in bscl cell, puri...

genetic typing methods of t. gondii strains have been extensively perfected in recent years. from a technical point of view, many tools usable for genetic studied on single-copy loci have been used: rflp, pcr-rflp, sequencing, rapd-pcr and isoenzyme analysis. we described the cloning and sequence analysis of the gene which encodes the major surface antigen (sag1 or p30) of t. gondii. sag1 is th...

Large-scale production of plasmid DNA to prepare therapeutic gene vectors or DNA-based vaccines requires a suitable bacterial host, which can stably maintain the plasmid DNA during industrial cultivation. Plasmid loss during bacterial cell divisions and structural changes in the plasmid DNA can dramatically reduce the yield of the desired recombinant plasmid DNA. While generating an HIV-based g...

AIM:To clone core gene cDNA of Chinese hepatitis C virus (HCV) into eukaryotic expression vector cosmid pTM3 and to express HCV core antigen in HepG2 cells.METHODS:Core gene cDNA of HCV was introduced into eukaryotic expression vector cosmid pTM3.Using vaccinia virus/bacteriophage T7 hybrid expression system, HepG2 cells were transfected with the recombinant plasmid pTM3-Q534 by lipofectin.RESU...

A recombinant E. coli, expressing nitrilase from Acidovorax facilis 72W with dual-site expression plasmid pRSFduet (E. coli pRSF-AfNit2), was constructed. It showed higher soluble of than that in the pET21a plasmid. The can efficiently catalyze hydrolysis 3-cyanopyridine to nicotinic acid. whole cells pRSF-AfNit2 were immobilized by using sodium alginate/glutaraldehyde/polyethylene imine as bes...

In order to examine new ideas for gene therapy in ovarian cancer, the specific mechanism underlying the effects of the WW domain containing oxidoreductase (WWOX) gene on cell cycle regulation and apoptosis in human ovarian cancer stem cells was investigated. Ovarian cancer stem cells were transfected with a eukaryotic expression vector carrying the WWOX gene in vitro (recombinant plasmid) and c...

The effects of the rolling-circle and theta modes of replication on the maintenance of recombinant plasmids in Lactococcus lactis were studied. Heterologous Escherichia coli or bacteriophage lambda DNA fragments of various sizes were inserted into vectors based on either the rolling-circle-type plasmid pWV01 or the theta-type plasmid pAMbeta1. All pAMbeta1 derivatives were stably maintained. pW...

the aim of this study was to produce a recombinant protein consisting of the catalytic and translocation domains of diphtheria toxin for its later application as a vaccine candidate against corynebacterium diphtheria. to achieve this goal, at first, the amino acid sequence of dt386 was used for prediction of t and b cell epitopes using on-line servers. the dt386 coding sequence was synthesized ...