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Introduction: Brucellosis is one of the most important zoonotic diseases that causes miscarriage and infertility in animals and causes human fever. The use of the common SS9 strain of Brucella abortus has several side effects for livestock. Brucella P39 protein is one of the plasma peripheral space proteins that is considered as one of the important immunogenic indicators. With the production of the new protein combination of P39, more studies can be done on the ability of this protein to stimulate immune responses against Brucella. Therefore, in this research, the production and purification of this protein in Escherichia coli bacteria has been done as a new compound.
method: In this experimental study, using the polymerase chain reaction, the P39 gene was propagated by the bacterium Brucella abortus. After purifying the P39 gene, it was cloned into plasmid carriers pSK+ and pGEX4T1. Therefore, pSK+-P39 and pGEX4T1-P39 structures were prepared. To produce the recombinant protein P39, the plasmid structure pGEX4T1-P39 first entered the Escherichia coli bacterium BL21. The protein was then produced by IPTG by induction of pGEX4T1-P39 plasmid. The resulting protein was purified using the orderly purification protein glutathione S-transferase. The amount of purified protein was measured using the Brad Ford method.
Results: The nucleotide sequence of the gene propagated by the cloned PCR in the plasmid carrier  pSK+ was exactly the same as the P39 gene of Brucella abortus. Production of P39 protein was performed by induction of pGEX4T1-P39 plasmid. The purified protein content was 200 micrograms per milliliter.
Conclusion: The production of the new protein P39 compound Brucella Abortus, which is unstable in the cytoplasm of the Escherichia coli bacterium, is possible using carriers with additive proteins such as pGEX4T1 in the host of Escherichia coli strain BL21.

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Introduction: In many studies, immunogenicity of Brucella proteins such as P39 in animals is investigated. In this study, we evaluated antigenicity of recombinant P39 from Brucella abortus in patients with Brucellosis. Materials and Methods: In this experimental study, at first recombinant P39 was produced in Escherichia coli. Sera reactivity of six infected individuals against the recombinant P39 protein was analysed by Western Blot. Results: Data indicated that P39 protein from Brucella abortus was recognized by patients, sera antibodies. Conclusion: Our data showed that recombinant P39 protein can be detected as an antigen by sera in infected human. Therefore, recombinant P39 have same epitopes with natural form of this antigen.

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Background: In molecular diagnosis of microbial agent, purification of chromosome is very important step. In this study, after cell destruction, DNA replication was done by increasing the denaturation time, without DNA purification. Methods and Materials: In this experimental study eight different dilution of E.coli (8/100, 4/100, 2/100, 1/100, 1/200, 1/400, 1/800 and 1/1600) solution were madce in D.W, Bacteria were separated by filtration. Polymerase chain reaction method was used to propagate 162 rRNA gene by design primers without DNA Purification. In order to confirme sensitivity of PCR, contamination of 15 different sources of Arak well water wafer was compared by MPN method. For confirmed sensitivity of PCR, 15 sources of water in Arak were examined and compared with MPN method. Results: Present of bacteria in diution sou tion were confirmed by culture. Polymerale Chain reaction (PCR) data were shown this method is able to recognize bacteria in above dilutions after filtration. This study showed high sensitivity of PCR method in compare to MPN method. Conclusion: Results were shown without stages of extraction of DNA, PCR were done without losing chromosome. Therefore false negative results were decrease and avoided from difficult phases.

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Background: Brucella is a gram-negative intracellular bacterium. Since Brucella brings about health and socio-economic problems, its control is of primary importance. The common method of vaccination includes using live attenuated strains of this bacterium. This study was done to evaluate the immunogenicity of Brucella aburtus P39 gene in Balb/c mice. Materials and Methods: In this experimental study, P39 gene was amplified by polymerase chain reaction (PCR) method and after extraction, it was sub-cloned to eukaryotic expression vector pcDNA3. The intramuscular injection of the obtained plasmid to the Balb/c mice was done in three stages. After the last vaccination, immunologic tests, such as proliferative response in lymphocytes, IFN- assessment, IL-5, and determination of IgG2a and IgG1, were run. Results: The level of activation in splenic lymphocytes response was 3.6 and the measured IFN- was 3 ng/ml, whereas IL-5 was insignificant. IgG2a and IgG1 titers were 1.640 and 1.40, respectively. Conclusion: The findings of the immunological analysis show the appropriate immune response in Balb/c mice model after the injection of P39 gene containing plasmid. The immune system response was in Th1 form which decreased the number of bacteria in spleen. Therefore, P39 gene is of appropriate immunogenicity and DNA vaccination is efficient in the activation of cell immune response against this bacterium.