Investigation of two Shuttle Vectors for Cloning and Expression of IL-11 Gene in Bacillus subtilis

Subject Areas : Microbial Biotechnology

َAtena Azarnosh ¹ (Department of Microbiology, Jahrom Branch)
majid moghbeli ² (Islamic Azad University, Damghan, Iran)
Farshid Kafilzadeh ³ (Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Mohammad Kargar ⁴ (Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Houshang Jamali ⁵ (Saatwinkler damm 147)

Received: 2020-06-28 Accepted : 2020-12-05 Published : 2020-12-21

Keywords: shuttle vector, Bacillus subtilis, pHT43, pMR12, interleukin-11,

Abstract :

Background & Objectives: The recombinant human IL-11 is the only approved medicine used for treating chemotherapy-induced side effects. Platelet count decreases (thrombocytopenia) in cancer patients who undergo chemotherapy. Interleukin-11 (IL-11) is a platelet increasing cytokine. This study aimed to use two shuttle vectors of pHT43 and pMR12 for cloning the IL-11 gene and     expression of its protein in Bacillus. subtilis and the expression level in these two vectors was   investigated.Materials & Methods: In this study, the IL-11 gene was designed and synthesized as a closed structure with two restriction sites for BamH1 and XbaI enzymes and a final length of 609 bp. Then the gene was cloned in two shuttle vectors of pHT43 and pMR12 and transferred to            B. subtilis WB 600. The expression level of the recombinant IL-11 was evaluated with the      Bradford incubation, the pMR12-int11-carrying bacteria expressed higher levels of the protein          (75 µg/mL) than pHT43-int11-carrying bacteria.Results: The results of PCR indicated that the IL-11 gene existed in shuttle vectors pHT43 and pMR12. The expression of this protein was about 75 /g / mL using pMR12 vector, which is higher than the bacterium carrying pHT43-int11.Conclusion: The expression level of the recombinant IL-11 protein with pMR12 vector was higher than pHT43 vector. This amount has not been reported so far for IL-11. B. subtilis can express and produce IL-11 and can be used as a source for drug production

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