Expression Of Fum5 Gene In E.Coli Strain Bl 21

Shruti, Yadav (2006) Expression Of Fum5 Gene In E.Coli Strain Bl 21. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type:	Student Project Report
Additional Information:	The objective of my project was to express the FUM5 gene (a polyketide Synthase gene required for fumonisin biosynthesis) in BL21 strain of E.coli and then purify the protein obtained by this expression. Fumonisins have been shown to be involved in many types of cancer in human being and animals, correlated with the consumption of Fumonisins. Fumonisins are produced by number of species with in Giberella fujikuroi species complex (Desjardin and Proctor, 1998). G.fujikuroi mP-A had been has been shown to be one of the most common pathogen of maize and found to be associated with disease of roots, stalk and ears. This fungus has shown to be present in apparently healthy maize tissue (Munkvold and Desjardins, 1997).Nothing is known about the molecular genetics of Fumonisin production. Classical genetic studies with natural variants and laboratory mutants of G.fujikuroi mP-A have identified four linked loci fum1, fum2, fum3 and fum4 that are involved in Fumonisin biosynthesis (Proctor,1999). Fum5 has been observed to be involved in the production of polyketide synthase (Proctor,1999) and have explored the involvement of isolated fum5 gene in the production of polyketide synthase. Other aspects considered include: preparation of competent cells of E.coli and their transformation using fum5 DNA and these transformants were selected on LB agar plates containing Ampicillin so that only resistant variety could grow; Presence of Plasmid to be checked by isolating it by SDS alkaline denaturation method and restriction digestion of recombinant plasmid for insert release; Transformants to be grown on induction medium containing optimized concentration of IPTG and then perform SDS PAGE to observe the expression of protein; Elution of expressed protein.
Uncontrolled Keywords:	E.coli FUM5 gene
Subjects:	600 Technology > 05 Chemical engineering > 04 Fermentation Technology
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	06 Mar 2007
Last Modified:	28 Dec 2011 09:26
URI:	http://ir.cftri.res.in/id/eprint/446

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