Background: The purpose of this study was to clone, express, and purify a novel multidomain fusion protein of Micobacterium tuberculosis (Mtb) in a prokaryotic system. Methods: An hspX/esxS gene construct was synthesized and ligated into a pGH plasmid, E. coli TOP10 cells were transformed, and the vector was purified. The vector containing the construct and pET-21b (+) plasmid were digested ...

The levels of intramolecular plasmid recombination, following transfection of a plasmid substrate for homologous recombination into normal and immortally transformed cells, have been examined by two independent assays. In the first assay, recovered plasmid was tested for DNA rearrangements which regenerate a functional neomycin resistance gene from two overlapping fragments. Following transform...

The incidence of pancreatic carcinoma, a gastrointestinal malignancy, is on the increase and effective therapeutic strategies are therefore required. This study aimed to construct a recombinant plasmid pcDNA3.1(-) shCEACAM6-yCDglyTK from CEACAM6 targeting shRNA and the fusion suicide gene yCDglyTK for inhibition of SW1990 human pancreatic carcinoma cell growth and invasion. A plasmid containing...

conclusions the gp40/15 gene, which was cloned in the pet28a+, was successfully expressed and produced in e. coli. therefore, this protein can be used in future studies to develop recombinant vaccines and diagnostic kits. methods in this experimental study, the gp40/15 gene sequence was extracted from genbank (no. af155624) and cloned in the pet28a+ plasmid. colony polymerase chain reaction (pc...

Background: Outer inflammatory protein A (OipA) is one of the important adhesins of H. pylori and a valuable candidate for vaccine development. Its gene is under "on-off" switch status which correlates with OipA protein expression. Objectives: We aimed to obtain a recombinant OipA clone (with "on" status) from an Iranian clinical isolate. Materials and Methods: A clinical H. pylori-isolate demo...

the identification of a large number of antigens with potential for development of new tuberculosis vaccine has been accomplished in recent years. this study was designed for cloning and expression of esat-6 as a potent antigen of mycobacterium tuberculosis. selected gene (rv3875) was amplified by pcr and product was ligated into expressing plasmid vector pqe30 and recombinant pqe30-es plasmid ...

results cloning and subcloning of the flic gene were confirmed by colony pcr, restriction enzymes digestion and dna sequencing of the recombinant plasmids pprime-flic and pvax-flic. the expression of flagellin protein in eukaryotic cells was approved by immunofluorescence assay (ifa), western blotting analysis and the reverse transcriptase polymerase chain reaction (rt-pcr) method. conclusions t...

DMSO Inoue transformation buffer (please see Step 1) Chilled to 0°C before use. Plasmid DNA (recombinant plasmid) Construct using one of the methods described in Directional Cloning into Plasmid Vectors, Attaching Adaptors to Protruding Termini, Blunt-ended Cloning into Plasmid Vectors, Dephosphorylation of Plasmid DNA, Addition of Synthetic Linkers to Blunt-ended DNAand Ligating Plasmid and Ta...

To clone the Eg95 and EgA31 antigen genes into the prokaryotic expression plasmid pET30a-EgA31-Eg95, we expressed the recombinant protein EgA31-Eg95 and confirmed with western blot analysis. The total RNA was extracted from the protoscoleces of Echinococcus granulosus (E. granulosus) adult worms. The complementary DNA (cDNA) encoding the EgA31 antigen was amplified via quantitative real-time po...

The protective antigen (PA) of Bacillus anthracis and the V antigen of Yersinia pestis are potent immunogens and candidate vaccine sub-units. When plasmid DNA encoding either PA or V antigen was used to immunise the Balb/c mouse, a low serum IgG titre was detected (log (10)1.0 or less) which was slightly increased by boosting with plasmid DNA. However, when mice immunised with plasmid DNA were ...