---

Background: Type III Interferon (IFN) is a novel member of the interferon family, which contains three ligands: IFN-λ1 (IL-29), IFN-λ2 (IL-28A) and IFN-λ3 (IL-28B).These three ligands use the same unique heterodimeric receptor composed of CRF2-12 (IFN-λ-R1/IL-28Ra) and CRF2-4 (IL10-R-b) chains which are completely different from type I & type II IFN receptors. IFNsλ exhibit several features such as antiviral activity, antiproliferative activity, immunomodulatory activity and in vivo antitumour activity. In this work we aimed to clone the ogene of IFN-λ1 obtained from dendritic cells and assess protein production in eukaryotic expression vector. Materials and methods: in thid experimental study, total RNA was extracted from monocyte derived dendritic cells stimulated with 100 ng/ml of LPS. cDNA was synthesized from total RNA .Then cDNA of IFN-λ1 was amplified by PCR with specific primers and cloned into the PTZ57R/Tvector in the E.coli (DH5α). This was subsequently subcloned into plasmid pcDNA3.1+, using KpnI and BamHI restriction endonucleases. After tranfection into HEK293 T, expression of protein was tested by sandwich-ELISA method. Results: The DNA sequence of the insert was identical to the published sequences encoding IFN-λ1 in GeneBank. It was demonstrated that IFN-λ1 gene was markedly transcribed in transfected cells. Expression of IFN-λ1 in HEK293 T cells was confirmed by sandwich ELISA. Conclusion: Successful cloning and expression IFN-λ1 can be the first step for more production and further investigation about other activities of this cytokine and provides grounds for research on obtaining new therapeutic approaches for cancer, viral, autoimmune and allergic disease and designing more effective vaccines.

---

Background: Helicobacter pylori (H. pylori) is a gram negative bacilli that causes the stomach and duodenum diseases in human. An important virulence factor of H. pylori is a CagA gene that increases of colonization it in stomach epithelial cells and lead to inflammation and peptic ulcers. The aim of the present study was to production of recombinant protein containing highly antigenic region of CagA in E. coli.

Materials and Methods: In this experimental study, the antigenic region (1245 base pair) of CagA gene was detected by bioinformatics methods, proliferated by PCR method, digested by BamHI and XhoI restriction enzymes and cloned into pET32a plasmid and was expressed in the E. coli BL21 (DE3) pLysS with induced by IPTG. The expressed protein was purified with Ni-NTA kit and its antigenicity was studied by western blotting method.

Results: Data showed the successful cloning and expression of the target gene. Recombinant CagA protein purified by Ni-NTA kit and dialysis with concentration of 1.5 mg/ml. In western blotting, the produced protein was interacted with infected human and mice sera.

Conclusion: Results indicated that recombinant CagA protein (65 KDa) maintains its antigenicity, so could be used for serological diagnosis of H. pylori diseases and production of vaccine.

---

Background: Shigella dysenteriae one of the main causes of diarrhea in humans, but there is no vaccine against it. IpaD protein is one of the most important virulence factors in pathogenic shigella. The cloning N-terminal ipaD genes with ctB genes that have a role in adjuvant and carrier as recombinant vaccine can caused enhance the mucosal immune response.

Materials and Methods: Designing primers for genes ctB and ipaD were carried out based on the construction of gene cassettes, respectively. PCR reactions were performed to amplify the fragments and amplified fragments were cloned into pGEM-Teasy vector. Both the vector cut by restriction enzymes HindIII and XhoI and ipaD gene to gene ctB finally were Fusion. The ctB-ipaD gene cassette and expression vector pET28a(+) cuted by SalI and HindIII restriction enzymes. The cloning ctB-ipaD cassette was performed in the expression vector and expression of gene cassettes.

Results: In this study, the N-terminal ipaD as vaccine candidate antigen was genetically linked to the C terminal of ctB which has a carrier and adjuvant role. Fusing ctB-ipaD in the expression vector pET28a(+) is confirmed by sequencing, PCR and digested with restriction enzymes. The recombinant proteins produced is confirmed by SDS-PAGE and Western blot.

Conclusion: According to previous and similar studies, product cassette ctB-ipaD and expression its was expected. Is hoped to protein expression of this gene cassette and the production of antibodies could be achieved the candidate vaccine against Shigella.

---

Background: Streptococcus pyogenes produce extracellular hyaluronidase enzyme which is directly associated with the spreading of the organism during infection. Hyaluronidase enzyme is able to break hyaluronic acid or interstitial cement. This enzyme might be used in cancer treatment.The objective of the present study was to clone and express the nucleotide sequence of this enzyme which is involved in hyaluronidase enzymatic activity.

Materials and Methods: The enzymatic region of hyaluronidase gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify the region. The amplified product was cloned into the expression vector pET32a. E. coli BL21 (DE3) pLYsS was transformed with recombinant plasmids. Then gene expression was induced by IPTG. The expressed protein was purified successfully via affinity chromatography by NiNTA kit. The integrity of the product was confirmed by western-blot analysis.

Results: The nucleotide sequence of amplified gene was consistent with the streptocuccal hyaluronidase gene. The concentration of recombinant protein calculated to 500 mg purified protein per liter. The enzymatic region of recombinant protein from Streptococcus pyogenes was recognized by all five patient’s sera with Streptococcus infection.

Conclusion: In general, it is possible to produce the enzymatic regions of the Streptococcus pyogenes hyaluronidase in Escherichia coli. The antigenic property of the produced protein is well retained. Considering the product's domestic demand and also low efficiency of production and pathogenicity of Streptococcus species, it is possible to produce it as recombinant product.