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The Creation of Neisseria gonorrhoeae Strains Constitutively Expressing Single Opa Proteins

Undergraduate #116
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology
Session: 2
Room: Exhibit Hall A

Princess Bush - Bennett College
Co-Author(s): Allison Palmer, University of Virginia, Charlottesville, VA; Alison K. Criss, University of Virginia, Charlottesville, VA



Neisseria gonorrhoeae (Gc) is the causative agent of the sexually transmitted infection gonorrhea. Neutrophils are recruited to the site of Gc infection but are unable to kill all the bacteria. Untreated Gc infection and neutrophilic inflammation can lead to serious complications such as pelvic inflammatory disease and infertility. Understanding how Gc interacts with neutrophils is important for finding new ways to treat the disease. The Gc outer surface membrane contains many outer membrane proteins, including opacity associated (Opa) proteins. Opas bind to carcinoembryonic antigen cell adhesion molecules (CEACAM), which are neutrophil receptors. Of most interest are CEACAMs 1 and 3, which lead to either inhibiting or activating signals within the neutrophils respectively when bound. Opa proteins can bind to all, some, or no CEACAMs, which may lead to different activation of the neutrophils after infection. This project seeks to create Gc strains that constitutively express a single Opa protein. We will complete this project by transforming Opa containing plasmids into a Gc strain with all its native Opa genes deleted, allowing us to look at one Opa specifically and investigate how it interacts with neutrophils. We can confirm successful transformation of the Gc by comparing it to our wild strain of Gc (negative control), Opa D expressing strain of Gc (positive control), and opaless strain of Gc (negative control). Our research indicates new challenges in Gc transformation in vitro. The creation of these Gc strains permits future research that can observe how this interaction affects Gc survival and neutrophil activation. This research could lead to findings that advance the current treatment for gonorrhea.

Funder Acknowledgement(s): This project was supported by R01 AI097312 from the NIH awarded to Alison Criss PhD, Associate Professor in the Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, VA. This program was supported by the NSF # 1712724 awarded to Willieta Gibson PhD, Interim Dean, Arts & Sciences/Associate Professor, Biology, Greensboro, NC.

Faculty Advisor: Dr. Alison Criss, akc2r@virginia.edu

Role: My role in this research project consisted of attempting to create three strains of Neisseria gonorrhoeae (Gc) that constitutively express a single opa protein. I was also able to test antibiotic concentrations for Gc transformation confirmation, purify plasmids, and purify genomic DNA for transformation prep. I was not able to transform a strain of Gc prior to the of my research internship.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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