One of the most basic techniques in biomedical research is cDNA cloning for expression studies in mammalian cells. Vaccinia topoisomerase I-mediated cloning (TOPO cloning by Invitrogen) allows fast and efficient recombination of PCR-amplified DNAs. Among TOPO vectors, a pcDNA3.1 directional cloning vector is particularly convenient, since it can be used for expression analysis immediately after...

introduction: helicobacter pylori (h. pylori) infection has remained as a global health problem. animal studies demonstrated the role of h. pylori oipa gene in the development of gastric cancer. the aim of this study was the cloning and expression of helicobacter pylori oipa gene in a bicistronic vector harboring mice il-18 gene. materials and methods: the target gene encoding oipa was amplifie...

due to the widespread applications of xylitol dehydrogenase, an enzyme used for the production of xylitol, the present study was designed for the cloning of xylitol dehydrogenase gene from glcunobacter oxydans dsm 2003. after extraction of genomic dna from this bacterium, xylitol dehydrogenase gene was replicated using polymerase chain reaction (pcr). the amplified product was entered into ptz5...

We have constructed two plasmid vectors for cloning and expression of DNA fragments controlled by the lac promoter as a NotI-expression cassette. Whereas plasmid pSJ33 allows mobilization of the expression cassette into a wide variety of Gram-negative bacteria by RP4-mediated conjugation, the low-copy-number plasmid pSJP18Not facilitates cloning and expression in Escherichia coli when high gene...

Background and Aims: DNA constructs containing HCV antigens have become one of the vaccine candidates for induction of anti-HCV cellular and humoral immunity. In this study, we constructed a novel expressing vector harboring a fusion sequence derived from an overlapping fragment in the middle of NS3 and a truncated core fragment to avoid troubles reported to be associated with full gene express...

results this study shows the prepared gene construct is inducible by glucose. gene expression of preproinsulin was observed in muscle tissue of rabbits. conclusions these finding indicated that research in diabetes mellitus gene therapy could be performed on larger animals. objectives the purpose of this study was to construct glucose inducible insulin gene plasmid and use it in the muscle tiss...

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: Interferon-gamma [IFN-γ) is the most important cytokine in immune system. This protein has been expressed bacterial cells. However, cloning not an easy task. We aimed to clone, express, and purify recombinant mouse IFN-γ overcome problems favor of commercial purposes. Materials Methods: To amplify gene product for cloning, we primarily designed two specific primers target gene. Foll...

brucellosis is a well-known domestic animal infectious disease, which is caused by brucella bacterium. the outer membrane protein 25 kda (omp25) gene plays an important role in simulating of tnf-α, ifn-α, macrophage, and cytokines cells. in the current study molecular cloning and expression analysis of omp25 gene for designing a subunit vaccine against brucella was investigated. amplifying the ...

Background and Aims: DNA cloning, sub-cloning and site directed mutagenesis are the most common strategies in nearly all projects of recombinant protein production. The classical method of restriction site cloning is unsatisfactory due to the need for supply of restriction enzymes and the inefficiency of the digestion reaction. Many new methods, including recombinatorial cloning and ligation in...