فهرست مطالب رضا جلالی راد

Influenza virus, which is a decisive agent of influenza, attracted interest because of the event of the annual epidemic among a wide range of animal and human hosts. This study aimed to express the hemagglutinin protein of Iranian swine Influenza A (H1N1) through a baculovirus system in SF9 cells to produce a new recombinant vaccine.

Hemagglutinin gene of Swine H1N1 virus was amplified with specific primers containing restriction enzymes site, and then cloned. Afterward, the vector was transformed to DH10Bac using Bac-to-Bac system in order to produce a recombinant bacmid. Hemagglutinin expression and its biological activity were assessed using molecular and immunization tests.

The target rHA in length, 1710 bp, was produced and expressed in transfected SF9 cells with a size of ~66 kDa. The infected cells expanded in size and their nucleus, and desiccated from the surface of the cell culture as a granular. They could absorb chick red blood cells (RBCs), and appear as cell aggregates forty-eight postinfection. The fact that infected cells were unable to form cell clamp showed the test's accuracy and inhibition of hemodesorption activity. The amount of protein obtained was 10.76 µg/100 µl, equal to 0.1 mg/ml.

The baculovirus expression system could express the recombinant protein in the insect cell. Therefore, it may be a well-suited alternative to produce a new generation of the vaccine instead of egg-based and cell-culturebased generation vaccines.

Cold plasma or non-thermal plasma technology is one of the methods of foodprocessing that is used to inactivate pathogen microorganisms and improve food safety. Coldplasma can affect the inactivation of a wide range of microorganisms, without harming thehost and healthy tissues.The aim of this study was to evaluate the effect of cold plasma directly on reducing thenumbers of Salmonella enteritidis on egg shell and also to determine the effect of plasmaexposure time and the composition of the injected gas.

In this study, the effect of cold atmospheric plasma radiation ofargon and argon containing 5% oxygen gas, at the rate of 3 liters / min, and at three differenttimes on inactivation of Salmonella enteritidis was investigated.

The effect of argon gas radiation for 1, 3 and 6 minutes on the reduction ofsalmonella enteritidis numbers being 4.490, 3.948, 0 cfu /ml respectively, was significant at1% probability level. Also, the radiation effect of argon containing 5% oxygen at 1, 3 and 6minutes,on reduction of Salmonella enteritidis numbers was 4.559, 4.226 and 0 cfu /ml,respectively. Both groups of the treatments caused a significant decreasing trend at thestatistical level of 1% as compared to the control sample.

The obtained data indicated that the effects of the gas type, as well as theirradiation time and their interaction on the tested bacterium were statistically significant atp>0/01 as compared to the control.

 The H1N1 influenza virus is a highly pathogenic virus that threatens human life. Vaccination is an effective way of preventing and controlling influenza. Production of recombinant hemagglutinin in the baculovirus expression system, in the insect eukaryotic cell substrate (Sf9), has been suggested as an effective strategy.

The H1N1 influenza virus hemagglutinin gene sequence was prepared from National Center for Biotechnology Information (NCBI). After designing a specific primer, the sequence was provided using restriction digestion, cloned into pFastBacHTA plasmid, and transferred to the DH10Bac cell to produce a recombinant bacmid. After extracting the relevant plasmid, it was transfused into the insect cell; and after the expression of the protein by Sf9 cell, the presence of recombinant protein was confirmed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot methods.

The hemagglutinin gene (654 bp) was cloned in pFastBacHTA plasmid using the two enzymes of BamHI and Xhol. Sf9 cell expressed a protein weighing approximately 60 kDa after receiving the recombinant bacmid protein. The extracted protein was identified and confirmed using SDS-PAGE and Western blot methods; and protein concentration was measured by Lowry method.

 The baculovirus system is useful for the production of proteins with complex structures. Generally, it can be concluded that this protein is highly expressed in insect cells. Due to the similarity of this system with the human system, it can be a suitable alternative for embryonic eggs in the future, and can be used in vaccination.