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Cloning of Glyphosate Degrading Gene in E. coli DH5α.

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Relation http://ir.cftri.com/11554/
 
Title Cloning of Glyphosate Degrading Gene in E. coli DH5α.
 
Creator Aastha, Mishra
 
Subject 26 Pesticide Chemistry
 
Description 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.
 
Contributor Manonmani, H. K.
 
Date 2013
 
Type Student Project Report
NonPeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://ir.cftri.com/11554/1/Aastha.pdf
Aastha, Mishra (2013) Cloning of Glyphosate Degrading Gene in E. coli DH5α. [Student Project Report] (Submitted)