In addition, results of nucleotide sequencing of the fragmented segment via PCR showed the presence of GAD gene. FINDINGS: Morphological, biochemical and genetic analyses of 16s rDNA sequencing indicated that the studied substrain was of the L. Plasmid pET32a-gad expression vector was transformed in Escherichia coli BL21 strain, and protein expression was assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. Afterwards, the gene was inserted into the pJET1.2/blunt cloning vector and subcloned in vector pET32a. Genomic DNA of the bacterium was isolated and amplified using the GAD gene via polymerase chain reaction (PCR. METHODS: In this experimental study, we investigated the morphological, biochemical, genetic and 16s rDNA sequencing of L. This study aimed to clone and construct the expression vector of GAD gene from Lactobacillus plantarum PTCC 1058 bacterium. Therefore, cloning of this enzyme is essential to the optimization of GABA production. GABA is synthesized by glutamic acid decarboxylase (GAD enzyme in many organisms, including bacteria. Obtained rOmp28 could be used as a research experimental tool to find its potential as a detection kit and vaccine candidate.Ĭloning and Expression Vector Construction of Glutamate Decarboxylase Gene from Lactobacillus Plantarumįull Text Available BACKGROUND AND OBJECTIVE: Gamma-aminobutyric acid (GABA is a four-carbon non-protein amino acid used in the treatment of hypertension, diabetes, inflammation, and depression. coli BL21 (DE3 and then recombinant protein was purified with Ni-NTA column of chromatography against His tag. Recombinant vector transformed and expressed in to E. melitensis bp26 gene was cloned first in to PTZ 57 R/T vector and accessed on the PET28a vector and sequenced. In Iran, Brucella melitensis is a common causative agent for brucellosis and BP26 protein of this bacterium having a good antigenesity and an important vaccine candidate. It is shown that although the vaccine can prevent abortion, it does not provide complete protection against infection. major mannose-1-phosphate guanyltransferase successfully.Īmplification, cloning and expression of Brucella melitensis bp26 gene (OMP28 isolated from Markazi province (Iran and purification of Bp26 Proteinįull Text Available Brucellosis is a debilitative disease that imposes costs on both economy and society. major mannose-1-phosphate guanyltransferase and others previously accepted in GenBank "nConclusion: We amplified and cloned Iranian L. There were some differences in amino acid sequences between Iranian L. PCR product was sequenced and deÂposited to GenBank. major mannose-1-phosphate guanyltransÂferase gene was extracted and confirmed by restriction analysis. "nResults: Recombinant plasmid containing 1140 bp as L. PCR product was cloned into pTZ 57 R and sub cloned into pET32a expression vector. DNA of Leishmania promastigotes was extracted and PCR reaction was done. major mannose-1-phosphate guanyltransferase sequence gene was deÂsigned and synthesized. In this study, we cloned mannose-1-phosphate guanyltransferase gene of Iranian L. In this way, development of candidate antigen for vaccine has speÂcial imÂportant. Gene Cloning of Iranian Leishmania major Mannose-1-Phosphate Guanyltransferaseĭirectory of Open Access Journals (Sweden)įull Text Available "nBackground: Leishmania is an obligatory intracellular protozoan parasite, which infects human beÂings when infected sand fly vector takes a blood meal. Most efforts are towards designing an effective vaccine to prevent leishmaniasis.
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