We optimized the expression and purification of outer membrane proteins SpaO LamB from Salmonella typhi. investigated various factors in processes, including use isopropyl β-d-1 thiogalactopyranoside (IPTG), imidazole, urea. First, PCR amplification was carried out on genes. The genes were then cloned pTZ57R/T, expressed pET28a vector transformed into Escherichia coli BL21 (DE3). Gene insertion...

The heterologous expression of the gene LACP83 (encoding a laccase) from Pleurotus ostreatus in Escherichia coli was characterized. The laccase enzyme activity and kinetics of bacterial growth with an inducer (IPTG) and without inducer were determined. The maximum enzymatic activity was observed at 7 h post induction with a value of 3740 ± 342 U/L, which was similar to that reported for the nat...

We fused obg, encoding an essential GTP-binding protein in Bacillus subtilis, to the LacI-repressible, IPTG (isopropyl-beta-D-thiogalactopyranoside)-inducible promoter Pspac. Depletion of Obg, following removal of IPTG, caused a defect in sporulation and in expression of sporulation genes that are activated by Spo0A approximately P. These defects were significantly relieved by a mutation in spo...

Here we describe a half-semester biochemistry laboratory course based on the expression, isolation, purification, and biochemical characterization of RecJ nuclease. The students transformed expression plasmid thermostable nuclease into host bacteria Escherichia coli BL21(DE3), to induce recombinant by IPTG. expressed was purified from cell lysates broken with sonication Ni-affinity chromatograp...

Oncogene activation and loss of tumor suppressor genes are known to play a role in tumor initiation as well as its progression.The potential roles of these genes in perturbation of genome stability has become of major interest. To better understand the relationship between expression of an oncogene and genetic instability, we have studied a cell line expressing an activated human Ha-ras under t...

Recombinantly produced enzymes are applied in many fields, ranging from medicine to food and nutrition, production of detergents, textile, leather, paper, pulp, plastics. Thus, the cost-effectiveness recombinant enzyme synthesis is an important issue biotechnological industry. Isopropyl-β-D-thiogalactoside (IPTG), analog lactose, currently most widely used chemical agent for induction synthesis...

BACKGROUND Besides Escherichia coli, Bacillus subtilis is an important bacterial species for the production of recombinant proteins. Recombinant genes are inserted into shuttle expression vectors which replicate in both E. coli and in B. subtilis. The ligation products are first transformed into E. coli cells, analyzed for correct insertions, and the correct recombinant plasmids are then transf...