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:: Volume 12, Issue 2 (11-2023) ::
gebsj 2023, 12(2): 251-261 Back to browse issues page
Cloning and expression of PVX coat protein in bacterium in order to produce carrier of anticancer drugs
Leila Pourhang , Maghsoud Pazhouhandeh * , Akbar Shirzad
Biotechnology Department, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran , pazhouhandeh@gmail.com
Abstract:   (454 Views)
The use of plant viruses as carriers of anti-cancer drugs in medicine is increasing day after day. One of the most important potato viruses is Potato X Potexvirus (PVX) which is globally expanded and is one of the first plant viruses used as a carrier of anticancer drugs. In this research, in order to produce PVX particles, its Coat protein (CP) gene was expressed. First, the RT-PCR reaction was performed for CP amplification using specific primers containing the endonuclease digestion sites. Electrophoresis of the PCR product on agarose gel confirmed the amplified fragment with a length of 714 bp. Then the CP gene and pGBKT7 plasmid were digested by BamHI and EcoRI enzymes and after purification, ligation was done between them. Recombinant plasmid pGBKT7-CPPVX was transferred to E. coli bacteria by electroporation. After selecting the bacteria containing the recombinant plasmid by PCR and extracting the plasmids from them, sequencing confirmed the correctness of the cloned gene sequence. Then, the CP gene was cloned into pET-21a expression plasmid by digestion and ligation method. The recombinant plasmid was transferred to BL21 strain of E. coli and cultured. By adding IPTG in the bacterial culture medium, CP protein was expressed from the plasmid and bacterial proteins were extracted. Protein electrophoresis showed the expression of CP. After purification, this protein can be used as a carrier of anti-cancer drugs.
Keywords: Cloning, Coat protein, drug carrier, PVX, protein expression
Full-Text [PDF 935 kb]   (145 Downloads)    
Type of Study: Research | Subject: Microrganisms and Viruses
Received: 2023/09/6 | Accepted: 2024/01/11 | Published: 2024/03/10
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Pourhang L, Pazhouhandeh M, Shirzad A. Cloning and expression of PVX coat protein in bacterium in order to produce carrier of anticancer drugs. gebsj 2023; 12 (2) :251-261
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Volume 12, Issue 2 (11-2023) Back to browse issues page
دوفصل نامه علمی-پژوهشی مهندسی ژنتیک و ایمنی زیستی Genetic Engineering and Biosafety Journal
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