Presentation Title

Molecular Cloning of the FabB and FabH enzymes in E. coli Bacteria

Presentation Type

Poster

Keywords

FabB, FabH, enzyme, molecular cloning, E. coli, antibiotics, drug discovery, PCR

Department

Chemistry

Major

Chemistry with emphasis in Biochemistry

Abstract

Genetic engineering is often used in the field of drug discovery to produce many copies of a specific enzyme in an attempt to discover possible inhibitors of the enzyme. The specific proteins FabH and FabB in E. coli are enzymes that serve as catalysts for fatty acid biosynthesis, a process essential for cell survival. Inhibiting specific steps in the fatty acid biosynthesis pathway has proven to be an effective strategy for developing antibacterial agents. Expression of FabH and FabB from plasmids in a molecular cloning experiment may be used for experiments to find possible inhibitors and antibacterial targets against E. coli bacteria. Using Polymerase Chain Reactions (PCR), a specific section of the isolated DNA for both FabB and FabH enzymes in E. coli were exponentially replicated. Plasmids containing the gene inserts were confirmed with a selective marker and amplified by insertion into an E. coli bacterial cell using heat shock and LiAc. The cells were replicated, and therefore the plasmid amplified, so that each cell had the same plasmid with the DNA of interest. Expression of the enzymes of interest was evaluated using gel electrophoresis.

Faculty Mentor

Dr. Matthew Joyner

Funding Source or Research Program

Not Identified

Location

Waves Cafeteria

Start Date

24-3-2017 2:00 PM

End Date

24-3-2017 3:00 PM

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Mar 24th, 2:00 PM Mar 24th, 3:00 PM

Molecular Cloning of the FabB and FabH enzymes in E. coli Bacteria

Waves Cafeteria

Genetic engineering is often used in the field of drug discovery to produce many copies of a specific enzyme in an attempt to discover possible inhibitors of the enzyme. The specific proteins FabH and FabB in E. coli are enzymes that serve as catalysts for fatty acid biosynthesis, a process essential for cell survival. Inhibiting specific steps in the fatty acid biosynthesis pathway has proven to be an effective strategy for developing antibacterial agents. Expression of FabH and FabB from plasmids in a molecular cloning experiment may be used for experiments to find possible inhibitors and antibacterial targets against E. coli bacteria. Using Polymerase Chain Reactions (PCR), a specific section of the isolated DNA for both FabB and FabH enzymes in E. coli were exponentially replicated. Plasmids containing the gene inserts were confirmed with a selective marker and amplified by insertion into an E. coli bacterial cell using heat shock and LiAc. The cells were replicated, and therefore the plasmid amplified, so that each cell had the same plasmid with the DNA of interest. Expression of the enzymes of interest was evaluated using gel electrophoresis.