An Improved Homologous Recombination Method for Rapid Cloning of the Antibody Heavy Chain Gene and Its Comparison with the Homologous Recombination and Traditional Cloning Methods

Background: The homologous recombination (HR) is one of the conventional cloning methods for the production of recombinant DNA. It is a quick method for in vivo DNA cloning without using the high cost restriction enzymes. A few modifications in the cloning procedure can increase the efficiency of this method.Materials and Methods: In this study, effect of heating on the rate of the IgG1 heavy chain gene cloning was investigated in the HR method and then it was compared with HR method without heating and traditional cloning method. For doing this comparison, three cloning methods including HR, HR with the heat treatment, and traditional cloning were used to clone the human IgG1 heavy chain into the pcDNA3.1(+) plasmid.Results: The results showed that adding heat in the HR method converts insert and vector from the double strand DNA to the single strand DNA. This allows them to copulate with each other better and faster than the two other methods. The heat addition also helps the cell enzyme system for a faster and easier recombination and moreover it increases the cloning efficiency of the HR method in case of in vitro processing.Conclusion: The results showed that giving heat in the HR method increases cloning rate 7.5% and this increase reaches 10% in comparison with the traditional method.

The association of interleukin-13 gene polymorphism withkala-azar patients

 Background& Objective: Host resistance towards Leishmania infection is mediated by cellular immune responses leading to macrophage activation and parasite killing. According to the important role of IL-13 in the defense against visceral leishmaniasis (VL) and the known effect of the IL-13 gene polymorphisms on its production, the aim of this study was to investigate the probable relationship between IL-13 gene polymorphisms and susceptibility to VL.   Materials & Methods: The patient group included 52 patients who had suffered from VL infection and the control group consisted of 104 non-relative healthy people from the same endemic areas the patients were from (southern part of Fars Province). IL-13 (position -1512 A/C) gene polymorphism was determined by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP).   Results: There was no significant association between the frequencies of IL-13 (-1512) alleles and genotypes in the patients with VL compared to the thenormal population.   Conclusion: This study indicated that the IL-13 (position -1512 A/C) genotypes cannot be considered as a genetic susceptibility factor for leishmaniasis.

Effect of Bacillus thuringiensis parasporal toxin on stimulating of IL-2 and IL-5 cytokines production

Introduction:Bacillus thuringiensis, is a Gram-positive spore-forming bacterium that produces crystalline parasporal protein (Cry) during sporulation. Some of these Cry toxins do not show cytotoxicity against insects but they are capable to kill some human and animal cancer cells. The aim of this study was to verify whether cytocidal parasporal of B thuringiensis strains have immunostimulatory activity on human peripheral blood mononuclear cells (PBMNC) and to evaluate the ability of IL-2 and IL-5 production. Materials and methods: B. thuringiensis toxin with cytocidal activity was isolated and treated with proteinase K. PBMNC was cultured and treated with activated crystal proteins. We evaluated the ability of different cytokines production with Flow Cytometry. Results: In this study, immune stimulatory toxins Cry1 were distinguished. This toxin can stimulate production of cytokines IL-2 and stop production of IL-5. Discussion and conclusion: According to anti-cancer effect of B. thuringiensis toxins and also immune stimulatory effect, with more research these toxins can be introduced as immunotherapy drug in cancer treatment

Relation between interleukin-13 gene polymorphisms and susceptibility to brucellosis in Iranian population

Brucella is an intracellular Gram-negative bacterium. Previous reports showed that gene polymorphisms of cytokines can affect resistance or susceptibility to Brucella infection. Interleukin-13, a cytokine secreted by Th2 lymphocytes, has an important role in immune responses against established infections. In this study, we investigated the association of three polymorphic sites of IL-13 with susceptibility to brucellosis in Iranian population. In this study 169 patients with brucellosis and 71 healthy controls were included. DNA was extracted and genotyped for three bi-allelic polymorphisms of IL-13 gene at positions -1512A/C, -1055C/T, and +2044G/A by polymerase chain reaction-restriction fragment length polymorphism method. None of the studied alleles and genotypes of IL-13 gene (-1512A/C, -1055C/T, and +2044G/A) showed significant relationship with susceptibility to brucellosis. However, among eight haplotypes, the distribution of TCG and CAA haplotypes were significantly higher in the patients compared with those in the controls (P=0.002 and P=0.034, respectively). Although, the later did not tolerate Bonferoni correction. On the contrary, the distribution of TCA haplotype was higher in the controls compared to that in the patients (P=0.01). Furthermore, TAG/TCA haplogenotypes were significantly higher among controls compared to the brucellosis patients (P=0.025). P value resulted from TCA and TAG/TCA did not tolerate Bonferroni correction. There is no association between the inheritance of different alleles and genotypes of interleukin-13 gene and susceptibility to brucellosis. However, it seems that the inheritance of some haplotypes and haplogenotypes of IL-13 can impact the susceptibility to brucellosis

Association of interleukin-2 gene variants (positions +114 and −384) and susceptibility to brucellosis in Iranian population

Brucella is an obligate intracellular gram negative bacterium and the causative agent of brucellosis. Interleukin-2 (IL-2) is a cytokine involved in cell-mediated immunity response secreted by activated T-cells and considered as the growth factor for T-cells. Previous reports have revealed that gene polymorphisms of cytokines can affect susceptibility to Brucella infection. The goal of this study was to investigate the relationship between IL-2 gene polymorphisms (positions +114 and −384) and susceptibility to brucellosis. A total of 173 brucellosis patients and 75 healthy animal husbandmen who had Brucella infected animals and consumed their contaminated dairy products, as control group, were included in this study. All participants were genotyped for IL-2 gene polymorphisms at positions +114 (G/T) and −384 (G/T), using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The frequency of TT genotype at the position +114 was significantly higher in the controls, compared with the patients. But, there was no significant difference between the groups regarding TG and GG genotypes as well as T and G alleles. Furthermore, at position −384, the frequencies of G allele and GG genotype were higher in the controls compared with patients, however, they were not significantly different Additionally, TT/TT haplogenotype (+114/−384) was significantly higher in the controls, compared with the patients. Conclusively, it is suggested that the inheritance of TT genotype (position +114) and TT/TT haplogenotype (+114/−384) of IL-2 gene could be considered as one of the genetic factors responsible for resistance to brucellosis

Putative secreted Lufaxin-like protein family from <i>Bichromomyia olmeca</i>.

<p>Putative secreted Lufaxin-like protein family from <i>Bichromomyia olmeca</i>.</p

Putative secreted Yellow salivary protein family from <i>Bichromomyia olmeca</i>.

<p>Putative secreted Yellow salivary protein family from <i>Bichromomyia olmeca</i>.</p