PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>

ABSTRACT Gene diversification is a common mechanism pathogens use to alter surface structures to aid in immune avoidance. Neisseria gonorrhoeae uses a gene conversion-based diversification system to alter the primary sequence of the gene encoding the major subunit of the pilus, pilE. Antigenic varia...

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Autores principales:	Egon A. Ozer, Lauren L. Prister, Shaohui Yin, Billy H. Ward, Stanimir Ivanov, H Steven Seifert
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	antigenic variation Pacific Biosciences sequencing amplicon sequencing gene diversification Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3c9ef33863f14e60b4c2a807535a0653
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spelling	oai:doaj.org-article:3c9ef33863f14e60b4c2a807535a06532021-11-15T15:27:32ZPacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>10.1128/mSphere.00562-192379-5042https://doaj.org/article/3c9ef33863f14e60b4c2a807535a06532019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00562-19https://doaj.org/toc/2379-5042ABSTRACT Gene diversification is a common mechanism pathogens use to alter surface structures to aid in immune avoidance. Neisseria gonorrhoeae uses a gene conversion-based diversification system to alter the primary sequence of the gene encoding the major subunit of the pilus, pilE. Antigenic variation occurs when one of the nonexpressed 19 silent copies donates part of its DNA sequence to pilE. We have developed a method using Pacific Biosciences (PacBio) amplicon sequencing and custom software to determine pilin antigenic variation frequencies. The program analyzes 37 variable regions across the strain FA1090 1-81-S2 pilE gene and can be modified to determine sequence variation from other starting pilE sequences or other diversity generation systems. Using this method, we measured pilin antigenic variation frequencies for various derivatives of strain FA1090 and showed we can also analyze pilin antigenic variation frequencies during macrophage infection. IMPORTANCE Diversity generation systems are used by many unicellular organism to provide subpopulations of cell with different properties that are available when needed. We have developed a method using the PacBio DNA sequencing technology and a custom computer program to analyze the pilin antigenic variation system of the organism that is the sole cause of the sexually transmitted infection, gonorrhea.Egon A. OzerLauren L. PristerShaohui YinBilly H. WardStanimir IvanovH Steven SeifertAmerican Society for Microbiologyarticleantigenic variationPacific Biosciences sequencingamplicon sequencinggene diversificationMicrobiologyQR1-502ENmSphere, Vol 4, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	antigenic variation Pacific Biosciences sequencing amplicon sequencing gene diversification Microbiology QR1-502
spellingShingle	antigenic variation Pacific Biosciences sequencing amplicon sequencing gene diversification Microbiology QR1-502 Egon A. Ozer Lauren L. Prister Shaohui Yin Billy H. Ward Stanimir Ivanov H Steven Seifert PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
description	ABSTRACT Gene diversification is a common mechanism pathogens use to alter surface structures to aid in immune avoidance. Neisseria gonorrhoeae uses a gene conversion-based diversification system to alter the primary sequence of the gene encoding the major subunit of the pilus, pilE. Antigenic variation occurs when one of the nonexpressed 19 silent copies donates part of its DNA sequence to pilE. We have developed a method using Pacific Biosciences (PacBio) amplicon sequencing and custom software to determine pilin antigenic variation frequencies. The program analyzes 37 variable regions across the strain FA1090 1-81-S2 pilE gene and can be modified to determine sequence variation from other starting pilE sequences or other diversity generation systems. Using this method, we measured pilin antigenic variation frequencies for various derivatives of strain FA1090 and showed we can also analyze pilin antigenic variation frequencies during macrophage infection. IMPORTANCE Diversity generation systems are used by many unicellular organism to provide subpopulations of cell with different properties that are available when needed. We have developed a method using the PacBio DNA sequencing technology and a custom computer program to analyze the pilin antigenic variation system of the organism that is the sole cause of the sexually transmitted infection, gonorrhea.
format	article
author	Egon A. Ozer Lauren L. Prister Shaohui Yin Billy H. Ward Stanimir Ivanov H Steven Seifert
author_facet	Egon A. Ozer Lauren L. Prister Shaohui Yin Billy H. Ward Stanimir Ivanov H Steven Seifert
author_sort	Egon A. Ozer
title	PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
title_short	PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
title_full	PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
title_fullStr	PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
title_full_unstemmed	PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
title_sort	pacbio amplicon sequencing method to measure pilin antigenic variation frequencies of <named-content content-type="genus-species">neisseria gonorrhoeae</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/3c9ef33863f14e60b4c2a807535a0653
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PacBio Amplicon Sequencing Method To Measure Pilin Antigenic Variation Frequencies of <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>

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