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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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Background: In recent years Influenza viruses have caused widely spread moderate to severe infection in all around the world and there is no Influenza vaccine which can protect people only with one dose injection till now. Therefore , producing a universal vaccine based on virus like particle (VLP) could be ideal. In this study one of the molecular structures was considered for VLP based Influenza vaccine. Materials and Methods: In this experimental study, the human influenza virus (A /New Caledonia 20/1999/ (H1N1)) was propagated in MDCK cell culture. Viral RNA was extracted using RNX-plus solution. Complementary DNA synthesis was carried out using uni-12 primer and random hexamer as specific and general primers, respectively. Neuraminidase open reading frame (1413-bp) was amplified by PCR and cloned into pBlue-script SK. Neuraminidase coding frame sub cloned into pFastBac11 plasmid through SalI/XhoI sites. After verification of cloned Neuraminidase by restriction analysis, it was subjected to automated sequencing bi-directionally. The recombinant pFastBac Neuraminidase vector was transformed to E.coli DH10Bac cells which harbor bacmid DNA and helper plasmid to create Neuraminidase recombinant bacmid. Results: Neuraminidase recombinant bacmid was created by homologous recombination between pFastBacNA and bacmid and was verified by PCR using Neuraminidase specific and M13 universal primers. Conclusion: Recombinant baculovirus expressing Neuraminidase gene can be also used with other individual recombinant baculoviruses expressing HA and M1 genes in production of influenza VLPs or proteins resulting from this structure could be purified in specific insects for vaccine research studies.

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Background: Genetic variability of influenza viruses causes new epidemics worldwide annually. Development of a new vaccine for prophylaxis of influenza virus has been amajor objective in recent years. The aim of this study was to construct a recombinant baculoviruscapable of expressing the two surficial antigenic glycoproteins, hemagglutininand neuraminidase, as well as matrix proteinsof swine influenza (H1N1) simultaneously and independently. Materials and Methods: In this experimental study, first, a triplet cassette providing simultaneous and independent expression of target proteins was designed and subjected to synthesis. It was then cloned into pFastBac1 donor plasmid. Competent E.ColiDH10Bac cells were transformed by donor clone and the recombinant bacmids were produced following homologous transposition. This construction was verified by PCR and then transfected into Sf9 insect cells to package new recombinant baculoviruses. Results: Restriction map of pFastBacI HNM1 donor plasmid confirmed the fidelity of the clone. The results of PCR done on the recombinant bacmidas template indicated that a proper homologous recombination has occurred between pFastBacI HNM1 donor plasmid and the bacmid in E.ColiDH10Bac host cells. Protein analysis of the infected Sf9 cells showed that all target proteins were efficiently expressed at the same time. Conclusion: The recombinant baculovirus constructed in this studypossesses proper characteristics to produce swine influenza virus-like particles in Sf9 cells.