Cloning of Glyphosate Degrading Gene in E. coli DH5α.

Aastha, Mishra (2013) Cloning of Glyphosate Degrading Gene in E. coli DH5α. [Student Project Report] (Submitted)

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Abstract

Glyphosate [N- (phosphonomethyl) glycine] is a broad spectrum, non-selective, systemic, post emergence herbicide used in the field of agriculture as well as domestically. It is the member of the amino acid herbicide family and its mode of action is through inhibition of 5-enolpyruvylskimate 3 phosphate synthase (EPSP), an enzyme of the shikimic pathway. The enzyme is important in the biosynthesis of aromatic amino acids phenylalanine, tyrosine and tryptophan. Glyphosate is being used world-wide in order to increase the crop production yield to feed the increasing population. It stimulates the central nervous system and toxic when fed in excess and is even mutagenic. Excessive exposure of glyphosate results in a number of health problems like irritation of the mouth, nausea, intestinal discomfort, vomiting, skin infections and diarrhoea. A large amount of Glyphosate in food is known to cause hypotension and pulmonoryedema. Excess of glyphosate is also reported to cause mutation, brain, intestinal and heart defects in foetuses, reduced head size, genetic alterations in the central nervous system, increase in the death of cells that help form the skull, and deformed cartilage. The adverse effects of glyphosate have warranted the development of several methods for removal of glyphosate from the soil. Conventional methods of degradation usually involve the processes such as chemical remediation, photodegredation and phytoremidiation. However, these methods suffer from drawbacks such as these processes are slow and require more time to degrade since glyphosate molecules has a high soil binding capacity which reduces the glyphosate degradation rate. In this report we present the cloning and sequencing of glyphosate degrading gene. Ten bacterial strains were screened for glyphosate degrading gene. The isolate Pseudomonas fluorescens biovar 1 gave the best PCR amplification. The glyphosate degrading gene was isolated by preparative agarose gel electrophoresis of PCR amplified product, cloned in to E.coli DH5α and partially sequenced. The sequence obtained was further studied for structural prediction and modelling of C-P lyase enzyme was done using bioinformatics tools.

Item Type:	Student Project Report
Uncontrolled Keywords:	Glyphosate N- (phosphonomethyl) glycine, herbicide, E.coli
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 26 Pesticide Chemistry
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	09 May 2014 09:41
Last Modified:	09 May 2014 09:41
URI:	http://ir.cftri.res.in/id/eprint/11554

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