{"id":241,"date":"2022-04-11T15:48:12","date_gmt":"2022-04-11T15:48:12","guid":{"rendered":"https:\/\/students.pingry.org\/irt\/?page_id=241"},"modified":"2023-03-31T12:38:46","modified_gmt":"2023-03-31T12:38:46","slug":"virf-2022-project","status":"publish","type":"page","link":"https:\/\/students.pingry.org\/irt\/virf-2022-project\/","title":{"rendered":"VirF – 2022\/3 Project"},"content":{"rendered":"

[et_pb_section fb_built=”1″ admin_label=”Hero” _builder_version=”4.4.7″ background_color=”#f8fbfe” use_background_color_gradient=”on” background_color_gradient_start=”#53a0fd” background_color_gradient_end=”#002960″ background_color_gradient_direction=”74deg” background_image=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2020\/07\/geometric-bg-overlay-01.jpg” background_blend=”overlay” custom_padding=”2.4%|0px||0px||” animation_style=”zoom” animation_direction=”bottom” animation_duration=”0ms” animation_intensity_zoom=”0%” animation_starting_opacity=”100%” box_shadow_style=”preset7″ box_shadow_horizontal=”0px” box_shadow_vertical=”-200px” box_shadow_color=”#F8FBFE”][et_pb_row _builder_version=”4.4.7″ custom_padding=”||0px|||”][et_pb_column type=”4_4″ _builder_version=”3.25″ custom_padding=”|||” custom_padding__hover=”|||”][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre||||||||” text_text_color=”#ffffff” text_font_size=”18px” text_line_height=”1.8em” header_font=”Nunito Sans|700|||||||” header_text_align=”center” header_text_color=”#ffffff” header_font_size=”45px” header_line_height=”1.3em” header_2_font=”||||||||” text_orientation=”center” max_width=”700px” module_alignment=”center” custom_margin=”||18px|||” custom_padding=”||17px|||” inline_fonts=”Abhaya Libre”]<\/p>\n

VirF 2022\/2023 Project –\u00a0Protein Purification of <\/b>Shigella dysenteriae <\/i><\/b>Transcription Factor VirF<\/b><\/span><\/h1>\n

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Maria Loss (Form VI), Maddie Humphreys (Form VI), Daniela Karnaugh (Form V), Katia Krishtopa (Form V)<\/span><\/h3>\n

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Project Summary + Project Goal\u00a0<\/h1>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][\/et_pb_section][et_pb_section fb_built=”1″ admin_label=”Features” _builder_version=”3.22″ background_color=”#f8fbfe” custom_margin=”-62px|||||” custom_padding=”151px|0px|12px|0px||”][et_pb_row _builder_version=”4.4.7″ min_height=”77px” custom_margin=”-10px|auto||auto||” custom_padding=”||0px|||”][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre||||||||” text_text_color=”#ffffff” text_line_height=”1.9em” background_color=”#cee1ff” width=”100%” max_width=”100%” min_height=”382px” max_height=”1000px” custom_margin=”-190px|-1px|0px|-2px|false|false” custom_padding=”14px||1px|4px|false|false” border_width_all=”5px” border_style_all=”inset”]<\/p>\n

Shigella is a pathogen that is a leading cause of bacterial food-borne illness worldwide. The virulence cascade of shigellosis requires one protein of the AraC superfamily transcription factors, VirF. Other members of this superfamily, such as ToxT which is involved in the virulence cascade of Vibrio cholerae, show inhibition of DNA binding by a small fatty acid. We wish to determine whether or not VirF can be inhibited in a similar manner, and therefore we are working to express, purify, and ultimately solve the crystal structures of VirF. Solving the structures will help to reveal the mechanism of transcriptional regulation in S. dysenteriae and could lead to development of novel treatments for Shigella related illnesses. The Shigella bacterium causes shigellosis, specifically mammalian gastroenteritis. An estimated 80-165 million cases of gastroenteritis due to Shigella species occur globally each year. Symptoms vary from inflammation of the intestines causing diarrhea to nausea and vomiting. Shigella also kills over 600,000 people annually. Infection begins with the ingestion of contaminated food or water. Many of the genes required for intestinal penetration, invasion of host cells, and intestinal and diarrheal disease are carried out by the major regulatory protein VirF located on the PINV as part of the regulatory cascade. The member of the AraC protein family is a DNA-binding protein that activates toxicity factors icsA and virB genes. These genes are then able to activate other downstream effectors involving invasion of the cell wall and host infection. Thus, inhibiting VirF would inhibit the whole virulence cascade of shigellosis.<\/span><\/h2>\n

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Current State<\/h2>\n

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Our team will be working at the laboratory at the Pingry School under the guidance of Dr. Jennifer Pousant and Dr. Morgan D\u2019Ausilio. We received the VirF plasmid dried on filter paper from Dartmouth\u2019s Kull Laboratory in January 2021. We rehydrated the plasmid in distilled water, and proceeded to conduct bacterial transformation to replicate the plasmid. Then, through a miniprep, we isolated the plasmid from the bacteria and sent the plasmid to a lab for sequencing. We then analyzed the sequencing\/plasmid map to confirm there are no mutations in our sample.<\/span><\/h2>\n

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Next Steps<\/span><\/h2>\n

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Our goal is to isolate the VirF proteins which will help to understand the mechanism of transcriptional regulation in S. dysenteriae. Understanding how to possibly inhibit these structures could lead to development of novel treatments for Shigella related illnesses. Our next steps are to run an autoinduction to express our target protein. We will then purify our protein through affinity chromatography. <\/span><\/h2>\n

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Research Week Poster\u00a0<\/span><\/h2>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][et_pb_row _builder_version=”4.4.7″ custom_margin=”|auto|-38px|auto||”][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/04\/For-Printing-VirF-Research-Week-Poster-2022.png” title_text=”For Printing – VirF Research Week Poster 2022″ _builder_version=”4.4.7″ custom_margin=”-39px|||||” border_width_all=”4px” border_style_all=”inset”][\/et_pb_image][\/et_pb_column][\/et_pb_row][\/et_pb_section][et_pb_section fb_built=”1″ admin_label=”Center Aligned Text ” _builder_version=”4.4.7″ background_color=”#f8fbfe” use_background_color_gradient=”on” background_color_gradient_start=”#53a0fd” background_color_gradient_end=”#002960″ background_color_gradient_direction=”74deg” background_image=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2020\/07\/geometric-bg-overlay-01.jpg” background_blend=”overlay” min_height=”223px” custom_margin=”28px||-281px|||” custom_padding=”0px|0px|0px|0px||” animation_direction=”bottom”][et_pb_row admin_label=”Tittle” _builder_version=”4.4.7″ custom_margin=”1px|auto||auto||”][et_pb_column type=”4_4″ _builder_version=”3.25″ custom_padding=”|||” custom_padding__hover=”|||”][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|700|||||||” header_text_align=”center” header_text_color=”#ffffff” header_font_size=”60px” custom_padding=”||0px|||”]<\/p>\n

Meet the Team<\/h1>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][et_pb_row column_structure=”1_4,1_4,1_4,1_4″ _builder_version=”4.4.7″ min_height=”85px” height=”87px” max_height=”100px” custom_margin=”||-2px||false|false” custom_padding=”0px||0px||false|false”][et_pb_column type=”1_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/04\/G11-Loss-M-1.TIF_.jpg” title_text=”G11 Loss M (1).TIF” _builder_version=”4.4.7″ min_height=”213px”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/04\/G11-Humphreys-M.TIF_.jpg” title_text=”G11 Humphreys M.TIF” _builder_version=”4.4.7″ height=”282px” custom_padding=”|0px|0px|||”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/04\/G10-Karnaugh-D.jpg” title_text=”G10 Karnaugh D” _builder_version=”4.4.7″ max_width=”97%” height=”282px” custom_padding=”0px||0px|||”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2023\/03\/zje1gp6ritzriagpkzzt.jpeg” title_text=”zje1gp6ritzriagpkzzt” _builder_version=”4.4.7″ hover_enabled=”0″][\/et_pb_image][\/et_pb_column][\/et_pb_row][et_pb_row column_structure=”1_6,1_6,1_6,1_6,1_6,1_6″ _builder_version=”4.4.7″ min_height=”23px” custom_margin=”|auto|-416px|auto||” custom_padding=”||16px|||”][et_pb_column type=”1_6″ _builder_version=”4.4.7″ custom_padding=”||0px||false|false”][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|600|||||||” header_text_align=”center” header_text_color=”#ffffff” header_2_font=”Abhaya Libre|600|||||||” header_2_text_color=”#ffffff” header_2_font_size=”35px” header_2_letter_spacing=”1px” width=”100%” custom_margin=”6px|||39px|false|false” custom_padding=”0px|0px|||false|false”]<\/p>\n

Maria Loss<\/h2>\n

[\/et_pb_text][\/et_pb_column][et_pb_column type=”1_6″ _builder_version=”4.4.7″][\/et_pb_column][et_pb_column type=”1_6″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|600|||||||” header_2_font=”Abhaya Libre|600|||||||” header_2_text_color=”#ffffff” header_2_font_size=”35px” width=”133.2%” custom_margin=”7px|||-70px|false|false” custom_padding=”|0px||||”]<\/p>\n

Maddie Humphreys<\/h2>\n

[\/et_pb_text][\/et_pb_column][et_pb_column type=”1_6″ _builder_version=”4.4.7″][\/et_pb_column][et_pb_column type=”1_6″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|600|||||||” header_2_font=”Abhaya Libre|600|||||||” header_2_text_color=”#ffffff” header_2_font_size=”35px” width=”124.2%” custom_margin=”6px|||-164px|false|false” custom_padding=”|0px||0px||”]<\/p>\n

Daniela Karnaugh<\/h2>\n

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Katia Krishtopa<\/h2>\n

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Dr. Jennifer Pousont<\/span><\/strong><\/h1>\n

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Dr. Morgan D’Ausilio<\/span><\/strong><\/h1>\n

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VirF 2022\/2023 Project –\u00a0Protein Purification of Shigella dysenteriae Transcription Factor VirFMaria Loss (Form VI), Maddie Humphreys (Form VI), Daniela Karnaugh (Form V), Katia Krishtopa (Form V)Project Summary + Project Goal\u00a0Shigella is a pathogen that is a leading cause of bacterial food-borne illness worldwide. The virulence cascade of shigellosis requires one protein of the AraC superfamily […]<\/p>\n","protected":false},"author":68,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_et_pb_use_builder":"on","_et_pb_old_content":"","_et_gb_content_width":"","footnotes":""},"class_list":["post-241","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/241","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/users\/68"}],"replies":[{"embeddable":true,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/comments?post=241"}],"version-history":[{"count":68,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/241\/revisions"}],"predecessor-version":[{"id":802,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/241\/revisions\/802"}],"wp:attachment":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/media?parent=241"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}