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Comparison of Tomato spotted wilt virus nucleocapsid gene expression in two different strains of E. coli
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Zahra Zaer   , Davoud Koolivand * , Omid Eini |
| Department of Plant Protection, Faculty of Agriculture, University of Zanjan, Iran , Koolivand@znu.ac.ir |
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Abstract: (5752 Views) |
Because of limitations in antibody production against plant viruses, coat protein expression has been developed to prepare antigen for antibody production. In the recent research, Nucleocapsid gene of Tomato spotted wilt virus was amplified from a local strain in Iran. Cloned segment in TA vector (pTG19TSWV-N) was sub-cloned into pET32a as expression vector that digested by XhoI and BamHI. Then, pET32TSWV-N was transformed in two different strains of E. coli BL21(DE3) and BL21(DE3) pLysS by heat shock method. For protein expression, the transformed cells were induced by 1mM of IPTG in both strains. The protein was extracted four hours after induction and analyzed in SDS-PAGE. The SDS-PAGE result showed that a 48 kDa protein have been expressed that is expected based on additional tags from plasmid. The result revealed that nucleocapsid had been expressed in both strains whereas protein expression was little more in BL21(DE3) pLysS.
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| Keywords: Expression, Prokaryotic expression, Recombinant protein. Tomato spotted wilt virus |
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Full-Text [PDF 923 kb]
(1801 Downloads)
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Type of Study: Research |
Subject:
Microrganisms and Viruses
Received: 2018/09/29 | Accepted: 2018/11/3 | Published: 2018/11/3
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