Presentation Title

Investigating the Growth Rate of a mntH Knockout Mutant of E. coli

Start Date

November 2016

End Date

November 2016

Location

HUB 302-36

Type of Presentation

Poster

Abstract

Under excessive amounts of hydrogen peroxide a cell will activate genes to protect itself against oxidation. In Escherichia coli, one of these genes, mntH, codes for a manganese import protein. Manganese is a redox cofactor and is thought to help protect proteins from oxidative damage. We wish to understand the role manganese may have in DNA damage and repair. To study this, we have attempted to knock out the mntH gene in a reporter strain of E. coli which is sensitive to DNA mutation. We used the Lambda Red system, a special recombination system, to insert a linear piece of DNA containing the kanamycin gene into the E. coli's genomic DNA, replacing the mntH gene. To optimize our technique, we successfully performed the knockout in a commercially available E. coli strain DH5-α, known to efficiently accept foreign DNA. We were able to generate a growth curve of DH5-alpha of the wild type and knockout mutant. Our results indicate that the mntH knockout grows more slowly than the wild type in the presence of hydrogen peroxide, which supports our hypothesis that mntH protects E. coli against oxidative stress. We are currently in the process of transforming the reporter strain CSH104 and our PCR insert and generating a growth curve for CSH104.

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Nov 12th, 1:00 PM Nov 12th, 2:00 PM

Investigating the Growth Rate of a mntH Knockout Mutant of E. coli

HUB 302-36

Under excessive amounts of hydrogen peroxide a cell will activate genes to protect itself against oxidation. In Escherichia coli, one of these genes, mntH, codes for a manganese import protein. Manganese is a redox cofactor and is thought to help protect proteins from oxidative damage. We wish to understand the role manganese may have in DNA damage and repair. To study this, we have attempted to knock out the mntH gene in a reporter strain of E. coli which is sensitive to DNA mutation. We used the Lambda Red system, a special recombination system, to insert a linear piece of DNA containing the kanamycin gene into the E. coli's genomic DNA, replacing the mntH gene. To optimize our technique, we successfully performed the knockout in a commercially available E. coli strain DH5-α, known to efficiently accept foreign DNA. We were able to generate a growth curve of DH5-alpha of the wild type and knockout mutant. Our results indicate that the mntH knockout grows more slowly than the wild type in the presence of hydrogen peroxide, which supports our hypothesis that mntH protects E. coli against oxidative stress. We are currently in the process of transforming the reporter strain CSH104 and our PCR insert and generating a growth curve for CSH104.