Presentation Title

Expression and purification of Listeria monocytogenes p60 protein

Start Date

November 2016

End Date

November 2016

Location

HUB 302-#14

Type of Presentation

Poster

Abstract

Listeria monocytogenes is a facultative anaerobic bacterium, which causes the infection listeriosis and is one of the most virulent foodborne pathogens. About 30% of food borne listeriosis infections in high-risk individuals such as pregnant patients and elderly individuals may be fatal. It is very important to develop rapid and effective methods for L. monocytogenes test to protect human health and avoid costly food recalls. Immunoassays are fast, reproducible and less sensitive to food interference, therefore are widely used in L. monocytogenes detection. However, the assays rely on the use of antibodies, and the binding of antibody to targets may not always be high and the quality of the antibodies is not consistent due to the nature of antibody production. Recently, aptamers emerged as a good alternative to antibody in diagnostics. Aptamers are short single-stranded oligonucleotides or peptides. Comparing to antibodies, they are more specific and stable. As they are produced by in vitro synthesis, their binding affinity is more consistent; therefore they are very suitable for use in L. monocytogenes test. In this study, we expressed and purified the extracellular protein p60 that is an ideal diagnostic target for the development of detection systems for L. monocytogenes. The GST-p60 fusion protein was expressed at high yield from E. coli and has been successfully purified to homogeneity. The recombinant protein will be used in the selection of specific binding DNA aptamers. The long-term goal of the project is to develop an aptamer-based fast and specific L. monocytogenes assay for use in food safety test.

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Expression and purification of Listeria monocytogenes p60 protein

HUB 302-#14

Listeria monocytogenes is a facultative anaerobic bacterium, which causes the infection listeriosis and is one of the most virulent foodborne pathogens. About 30% of food borne listeriosis infections in high-risk individuals such as pregnant patients and elderly individuals may be fatal. It is very important to develop rapid and effective methods for L. monocytogenes test to protect human health and avoid costly food recalls. Immunoassays are fast, reproducible and less sensitive to food interference, therefore are widely used in L. monocytogenes detection. However, the assays rely on the use of antibodies, and the binding of antibody to targets may not always be high and the quality of the antibodies is not consistent due to the nature of antibody production. Recently, aptamers emerged as a good alternative to antibody in diagnostics. Aptamers are short single-stranded oligonucleotides or peptides. Comparing to antibodies, they are more specific and stable. As they are produced by in vitro synthesis, their binding affinity is more consistent; therefore they are very suitable for use in L. monocytogenes test. In this study, we expressed and purified the extracellular protein p60 that is an ideal diagnostic target for the development of detection systems for L. monocytogenes. The GST-p60 fusion protein was expressed at high yield from E. coli and has been successfully purified to homogeneity. The recombinant protein will be used in the selection of specific binding DNA aptamers. The long-term goal of the project is to develop an aptamer-based fast and specific L. monocytogenes assay for use in food safety test.