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Paper Information

Journal:   GENETICS IN THE 3RD MILLENNIUM   WINTER 2016 , Volume 13 , Number 4; Page(s) 4112 To 4119.
 
Paper: 

CLONING, SEQUENCING, AND EXPRESSION OF FANC ANTIGEN OF ENTEROTOXIGENIC ESCHERICHIA COLI

 
 
Author(s):  NASIRI KHADIJEH, NASSIRI MOHAMMADREZA*, TAHMOORESPUR MOJTABA, HAGHPARAST ALIREZA, ZIBAEE SAEED
 
* DEPARTMENT OF ANIMAL SCIENCE AND INSTITUTE OF BIOTECHNOLOGY, FERDOWSI UNIVERSITY OF MASHHAD, MASHHAD, IRAN
 
Abstract: 

Enterotoxigenic Escherichia coli (ETEC) strains are the major cause of diarrhea in new born calves and certain species such as humans, pigs and sheep. ETEC has the ability to bind with entrocytes through K99 and F41, and to produce enterotoxins as a result. The purpose of this study was to produce the recombinant FanC protein of Enterotoxigenic Escherichia coli in prokaryotic system by using pET32a (+) expression vector. Therefore, using specific primers and polymerase chain reaction, FanC (K99) gene was amplified from Enterotoxigenic Escherichia coli. After purification, the gene was cloned and sub-cloned into pTZ57R/T and pET32a (+) respectively. The recombinant vector was transferred into E.oli BL21CodonPlus (DE3) as expression host, and the produced protein was purified by Ni-NTA chromatography column. The results of protein expression showed that E.coli was able to express FanC protein appropriately. This gene was expressed by the induction of IPTG at the concentration of 1mM. This was confirmed by Ni–NTA column, dot-blotting analysis and SDSPAGE electrophoresis. The concentration of the recombinant protein was calculated as 1.3 mg/ml.
The results of this study showed that E.coli can be used as an appropriate host to produce the recombinant FanC protein. This recombinant protein may be suitable to produce immunoglobulin, recombinant vaccine and diagnostic kit in future studies.

 
Keyword(s): ENTEROTOXIGENIC ESCHERICHIA COLI, CLONING MOLECULAR, GENE EXPRESSION, RECOMBINANT PROTEIN
 
 
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Click to Cite.
APA: Copy

NASIRI, K., & NASSIRI, M., & TAHMOORESPUR, M., & HAGHPARAST, A., & ZIBAEE, S. (2016). CLONING, SEQUENCING, AND EXPRESSION OF FANC ANTIGEN OF ENTEROTOXIGENIC ESCHERICHIA COLI. GENETICS IN THE 3RD MILLENNIUM, 13(4), 4112-4119. https://www.sid.ir/en/journal/ViewPaper.aspx?id=508656



Vancouver: Copy

NASIRI KHADIJEH, NASSIRI MOHAMMADREZA, TAHMOORESPUR MOJTABA, HAGHPARAST ALIREZA, ZIBAEE SAEED. CLONING, SEQUENCING, AND EXPRESSION OF FANC ANTIGEN OF ENTEROTOXIGENIC ESCHERICHIA COLI. GENETICS IN THE 3RD MILLENNIUM. 2016 [cited 2021May13];13(4):4112-4119. Available from: https://www.sid.ir/en/journal/ViewPaper.aspx?id=508656



IEEE: Copy

NASIRI, K., NASSIRI, M., TAHMOORESPUR, M., HAGHPARAST, A., ZIBAEE, S., 2016. CLONING, SEQUENCING, AND EXPRESSION OF FANC ANTIGEN OF ENTEROTOXIGENIC ESCHERICHIA COLI. GENETICS IN THE 3RD MILLENNIUM, [online] 13(4), pp.4112-4119. Available: https://www.sid.ir/en/journal/ViewPaper.aspx?id=508656.



 
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