AMPLIFICATION, CLONING AND EXPRESSION ASSAY OF THE MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE) FROM ENTEROTOXIGENICESCHERICHIA COLI (ETEC)

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

MANSOURI M.; MOUSAVI S.J.; NAZARIAN SH.; EHSAEE Z.; JAFARI F.; KHALESI R.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: Journal of Molecular and Cellular Researches Year:2013 | Volume:26 | Issue:2 Page(s): 221-228

Download Full-Text

Persian Verion

View:

949

Download:

Cites:

Information Journal Paper

Title

AMPLIFICATION, CLONING AND EXPRESSION ASSAY OF THE MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE) FROM ENTEROTOXIGENICESCHERICHIA COLI (ETEC)

Author(s)

Keywords

ENTEROTOXIGENIC ESCHERICHIA COLI (ETEC)Q2

MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE)Q2

CLONINGQ1

RECOMBINANT PROTEIN EXPRESSIONQ2

Abstract

Enterotoxigenic Escherichia coli (ETEC) is the most common cause of bacterial diarrhea in children under 5 years and it is also a cause of Travelers diarrhea worldwide. To prevent ETEC-caused diarrhea and decrease its prevalence, the WHO has promoted the vaccine production against this pathovar. CFA/I fimbriae is an important and frequent virulence factor of this bacteria, playing a critical role in pathogenesis. Therefore, tip protein of this fimbriae (CFaE) could describe as a vaccine candidate. This study was aimed at investigating the expression of the gene rCFaE from native sequence after CLONING into expression vector. rCFaE was amplified by PCR using a newly designed set of primers. PCR product was then cloned into early CLONING vector pTZ57R/T and then sub cloned into the expression vector pET28a(+). Protein expression in 2 strains of E.coli BL21-(DE3) pLysS and Rosetta was determined by using IPTG under different conditions. CLONING and sub CLONING process were performed successfully. Test samples in the comparatives with control samples did not show detectable protein on the SDS-PAGE. The same results were obtained after changing different parameters like time and temperature of induction and variety of IPTG concentration. It was not possible to obtain high level expression of the target recombinant protein production in E.coli with pattern codon usages, probably due to the high A+T content and also abundance rare codons in the native sequence of cfaE gene. Therefore we suggest synthesizing this gene after codon optimization and checking for expression that again.

Cites

No record.

References

No record.

Cite

APA: Copy

MANSOURI, M., MOUSAVI, S.J., NAZARIAN, SH., EHSAEE, Z., JAFARI, F., & KHALESI, R.. (2013). AMPLIFICATION, CLONING AND EXPRESSION ASSAY OF THE MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE) FROM ENTEROTOXIGENICESCHERICHIA COLI (ETEC). JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 26(2), 221-228. SID. https://sid.ir/paper/248585/en

Vancouver: Copy

MANSOURI M., MOUSAVI S.J., NAZARIAN SH., EHSAEE Z., JAFARI F., KHALESI R.. AMPLIFICATION, CLONING AND EXPRESSION ASSAY OF THE MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE) FROM ENTEROTOXIGENICESCHERICHIA COLI (ETEC). JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2013;26(2):221-228. Available from: https://sid.ir/paper/248585/en

IEEE: Copy

M. MANSOURI, S.J. MOUSAVI, SH. NAZARIAN, Z. EHSAEE, F. JAFARI, and R. KHALESI, “AMPLIFICATION, CLONING AND EXPRESSION ASSAY OF THE MINOR SUBUNIT COLONIZATION FACTOR ANTIGEN I (CFAE) FROM ENTEROTOXIGENICESCHERICHIA COLI (ETEC),” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 26, no. 2, pp. 221–228, 2013, [Online]. Available: https://sid.ir/paper/248585/en

Related Journal Papers