Subcloning and expression of SO6 gene, Saponaria Officinalis plant in
E.coli and investigation of antibody titer in rats

abdollahi, masoud; honari, hossein; nazarian, shahram; masoudi kerahroudi, mehdi

Volume 24, Issue 12 (Feb-Mar 2017) JSSU 2017, 24(12): 1024-1033 | Back to browse issues page

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abdollahi M, honari H, nazarian S, masoudi kerahroudi M. Subcloning and expression of SO6 gene, Saponaria Officinalis plant in E.coli and investigation of antibody titer in rats . JSSU 2017; 24 (12) :1024-1033
URL: http://jssu.ssu.ac.ir/article-1-3906-en.html

Subcloning and expression of SO6 gene, Saponaria Officinalis plant in E.coli and investigation of antibody titer in rats

Masoud Abdollahi

, Hossein Honari ^*

, Shahram Nazarian

, Mehdi Masoudi kerahroudi

Abstract: (5680 Views)

Introduction: Plant ribosome inactivating proteins act as N-glycosidase enzyme and produce by several family of Caryophyllaceae such as Saponaria Officinalis. Different Isoforms of RIPs expressed by Saponaria Officinalis. SO6 isoform depurinate Adenine 4324 in the conserved GAGA loop of 28SrRNA and disrupts protein synthesis. The aim of this study was expression of SO6 isoform in E.coli and investigation of antibody titer in rats.

Methods: In this experimental study, SO6 synthetic gene was excised from recombinant pUC57- SO6 plasmid with BamHI and SalI restriction enzymes and subcloned into pET28a (+) expression vector. The expression of recombinant protein was induced by IPTG. Recombinant SO6 was purified by nickel affinity chromatography. Western blotting was performed to confirm the recombinant protein. Rats were immunized intraperitoneal with purified protein and IgG serum titer was assayed by ELISA.

Results: PCR reaction and enzyme digestion confirmed subcloning of SO6 gene into pET28a (+) expression vector. A 29.5kDa protein band on SDS-PAGE showed a high level of recombinant protein expression. Polyclonal antibodies recognized SO6. ELISA confirmed significant antibody titer after injection of protein in test group compared with the control group.

Conclusion: The recombinant purified SO6 antigen can be used for anti-cancer and vaccine candidate research.

Keywords: N-glycosidase, Saponaria Officinalis, RIP, SO6

Full-Text [PDF 1182 kb] (2007 Downloads)

Type of Study: Original article | Subject: Genetics
Received: 2016/10/1 | Accepted: 2017/02/18 | Published: 2017/06/6

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