Event Title

Regulation of Cronobacter Sakazakii Apee Outer Membrane Esterase

Presenter Information

Oumar Sanogo, Minnesota State Univer

Location

CSU Ballroom

Start Date

16-4-2013 10:00 AM

End Date

16-4-2013 12:00 PM

Student's Major

Biological Sciences

Student's College

Science, Engineering and Technology

Mentor's Name

Christopher Conlin

Mentor's Department

Biological Sciences

Mentor's College

Science, Engineering and Technology

Description

C. sakazakii bacterial infection is mainly observed in infants, where the primary source of the organism and the main vehicle for its transmission is rehydrated, powdered infant formula. The organism is also found in many plant materials such as cereals, wheat, corn, soy, rice, herbs and spices; in rats, and flies; and in other food products. To begin studying the possible roles of the outer membrane esterase ApeE in this organism, we have cloned the promoter for the C.sakazakii apeE gene and begun studying its regulation. First, we amplified the promoter region of the C.sakazakii apeE gene outer membrane esterase through the polymerase chain reaction (PCR) process, and then cloned the PCR fragments into the pRS550 vector where it drives expression of the gene for the easily assayed enzyme galactosidase. Second, we transfered the pRS550+PCR fragment reporter plasmid into strains of E. coli containing deletions of various regulatory genes, and the amount of galactosidase expressed from the C. sakazakii apeE promoter was measured. Our results showed that galactosidase expression in the rpoS mutant was reduced by about 50 fold compared to the wild type. This is different from the regulation of the homologous gene in the related bacterium Salmonella enterica where it is regulated by PhoB. Since RpoS regulates genes that are highly expressed when the bacterium enters stationary phase, we intend to confirm these results by studying expression of the C. sakazakii apeE gene during both exponential growth and stationary phase.

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Apr 16th, 10:00 AM Apr 16th, 12:00 PM

Regulation of Cronobacter Sakazakii Apee Outer Membrane Esterase

CSU Ballroom

C. sakazakii bacterial infection is mainly observed in infants, where the primary source of the organism and the main vehicle for its transmission is rehydrated, powdered infant formula. The organism is also found in many plant materials such as cereals, wheat, corn, soy, rice, herbs and spices; in rats, and flies; and in other food products. To begin studying the possible roles of the outer membrane esterase ApeE in this organism, we have cloned the promoter for the C.sakazakii apeE gene and begun studying its regulation. First, we amplified the promoter region of the C.sakazakii apeE gene outer membrane esterase through the polymerase chain reaction (PCR) process, and then cloned the PCR fragments into the pRS550 vector where it drives expression of the gene for the easily assayed enzyme galactosidase. Second, we transfered the pRS550+PCR fragment reporter plasmid into strains of E. coli containing deletions of various regulatory genes, and the amount of galactosidase expressed from the C. sakazakii apeE promoter was measured. Our results showed that galactosidase expression in the rpoS mutant was reduced by about 50 fold compared to the wild type. This is different from the regulation of the homologous gene in the related bacterium Salmonella enterica where it is regulated by PhoB. Since RpoS regulates genes that are highly expressed when the bacterium enters stationary phase, we intend to confirm these results by studying expression of the C. sakazakii apeE gene during both exponential growth and stationary phase.

Recommended Citation

Sanogo, Oumar. "Regulation of Cronobacter Sakazakii Apee Outer Membrane Esterase." Undergraduate Research Symposium, Mankato, MN, April 16, 2013.
https://cornerstone.lib.mnsu.edu/urs/2013/poster-session-A/4