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Comparative study of transformation techniques of the Bacillus megaterium by eletroporation and protoplasts for recombinant protein production

Grant number: 12/21348-2
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2013
Effective date (End): December 31, 2013
Field of knowledge:Engineering - Chemical Engineering - Chemical Process Industries
Principal researcher:Rosineide Gomes da Silva Cruz
Grantee:Renata Porto Sampaio
Home Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

This project aims to continue the studies of transformation steps of Bacillus megaterium concerning recombinant protein production. The Biochemical Engineering research area in Chemical Engineering Department at Universidade Federal de São Carlos (ChE/UFSCar) has a molecular biology laboratory where studies of penicillin G acylase (PGA) production by Bacillus megaterium have been carried out. These studies aim both to the understanding of the process and the training of qualified personnel. The first studies were conducted employed a wild strain of Bacillus megaterium. The objective was to increase the expression of PGA. Then, studies concerning enzyme expression using recombinant microorganisms were carried out. One of the major difficulties found is in the Bacillus megaterium transformation stage. A bacterial transformation consists in inserting a vector, in this case, a plasmid (exogenous DNA), in a bacterium. This provides new characteristics to the bacterium, such as resistance to certain antibiotics and the ability to produce a molecule of commercial interest. The transformation is considered a critical stage and an event of central importance in the cloning process. The focus of this project is to continue the studies of the transformation techniques in B. megaterium, a gram-positive microorganism, as well as E. coli. Until this moment, there were no successful attempts of the microorganism electroporation with the protocols used. Thus, given the difficulties encountered in the transformation of Bacillus megaterium is that this project is inserted. It is hoped thereby to enhance the research in the cloning of recombinant protein gene and give the capability for an undergraduate student to work in laboratories of biochemistry and molecular biology, with the intent of enriching and completing his formation.(AU)

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