Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase
Glaesserella parasuis is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for G. parasuis yet. To est...
Gespeichert in:
| Veröffentlicht in: | Applied microbiology and biotechnology 2022-08, Vol.106 (13-16), p.5167-5178 |
|---|---|
| Hauptverfasser: | , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 5178 |
|---|---|
| container_issue | 13-16 |
| container_start_page | 5167 |
| container_title | Applied microbiology and biotechnology |
| container_volume | 106 |
| creator | Xiao, Jing Wang, Qiaochu Xiao, Kunxue Zhu, Wenlong Huang, Junhao Cai, Xuwang Chen, Huanchun Xu, Xiaojuan |
| description | Glaesserella parasuis
is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for
G. parasuis
yet. To establish a markerless knockout, deleted allelic genes with kanamycin resistance (Kan
R
) cassettes were introduced into the genome of
G. parasuis
by using natural transformation with suicide plasmids. Then, the Kan
R
cassette was excised with a thermosensitive plasmid pGF conferring a constitutive Flp expression. To realize the markerless and multiple-gene knockout, plasmid pGAF was constructed by placing the Flp gene under the control of an arabinose-inducible promoter. Firstly, pGAF was introduced into
G. parasuis
by electroporation, and the marked mutants were produced following natural transformation. Finally, the Kan
R
cassette was excised from the genome by the inducible expression of Flp upon arabinose action. Based on the natural transformation and the inducible expression of Flp, the markerless single-gene knockout mutants of Δ
hsdR
, Δ
neuA2
, Δ
espP2
, Δ
apd
, and Δ
nanH
were constructed. In addition, a five-gene knockout mutant of Δ
hsdR
Δ
neuA2
Δ
espP2
Δ
apd
Δ
nanH
was generated by successive natural transformation with five suicide plasmids. Taken together, a markerless and multiple-gene deletion system was established for
G. parasuis
in the present study for the first time. This system is simple, efficient, and easy to manipulate for
G. parasuis
; thus, our technique will substantially aid the understanding of the etiology, pathogenesis, and genetic engineering of
G. parasuis
and other bacteria that can be naturally transformed in laboratory conditions.
Key points
• Flp recombinase excised the Kan
R
gene flanked by FRT sites in
Glaesserella parasuis.
• The regulatory expression of Flp enabled a multiple-gene knockout for
G. parasuis.
• The technique will promote the understanding of Glässer’s disease pathogens.
Graphical abstract |
| doi_str_mv | 10.1007/s00253-022-11994-z |
| format | Article |
| fullrecord | <record><control><sourceid>gale_proqu</sourceid><recordid>TN_cdi_proquest_miscellaneous_2691788188</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A711762732</galeid><sourcerecordid>A711762732</sourcerecordid><originalsourceid>FETCH-LOGICAL-c453t-5f0ca3b2ac7e9ef6acadb98ca02fd4993e49f34bc9fff7cf1f654c1182a59bdc3</originalsourceid><addsrcrecordid>eNp9ks1rFTEUxQdR8Fn9B1wF3OhiapL5yGRZin0tFAQ_1uFO5uaRvkwyJjNgu_JPN9MRyhNxdSH8zrnnklMUbxk9Z5SKj4lS3lQl5bxkTMq6fHhW7Fhd8ZK2rH5e7CgTTSka2b0sXqV0RynjXdvuil979BhhtsGTYMgI8YjRYUoE_EDGxc12clgeMkWOPuhjWGZiPdk7yBBGdA7IBBHSYhPpIeFAspWHeYngyBzBJxPiuG1YPa_cRCLqMPbWZ_x18cKAS_jmzzwrvl99-nZ5Xd5-3t9cXtyWum6quWwM1VD1HLRAiaYFDUMvOw2Um6GWssJamqrutTTGCG2YaZtaM9ZxaGQ_6OqseL_5TjH8WDDNarRJr_E9hiUp3komuo51XUbf_YXehSX6nG6lGtY1teBP1AEcKutNyMfq1VRdCMZEy0W1Uuf_oNb0OFodPBqb308EH04EmZnx53yAJSV18_XLKcs3VseQUkSjpmjzF94rRtXaC7X1QuVeqMdeqIcsqjZRyrA_YHy67j-q30BnvPE</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2695185472</pqid></control><display><type>article</type><title>Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase</title><source>SpringerLink Journals - AutoHoldings</source><creator>Xiao, Jing ; Wang, Qiaochu ; Xiao, Kunxue ; Zhu, Wenlong ; Huang, Junhao ; Cai, Xuwang ; Chen, Huanchun ; Xu, Xiaojuan</creator><creatorcontrib>Xiao, Jing ; Wang, Qiaochu ; Xiao, Kunxue ; Zhu, Wenlong ; Huang, Junhao ; Cai, Xuwang ; Chen, Huanchun ; Xu, Xiaojuan</creatorcontrib><description>Glaesserella parasuis
is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for
G. parasuis
yet. To establish a markerless knockout, deleted allelic genes with kanamycin resistance (Kan
R
) cassettes were introduced into the genome of
G. parasuis
by using natural transformation with suicide plasmids. Then, the Kan
R
cassette was excised with a thermosensitive plasmid pGF conferring a constitutive Flp expression. To realize the markerless and multiple-gene knockout, plasmid pGAF was constructed by placing the Flp gene under the control of an arabinose-inducible promoter. Firstly, pGAF was introduced into
G. parasuis
by electroporation, and the marked mutants were produced following natural transformation. Finally, the Kan
R
cassette was excised from the genome by the inducible expression of Flp upon arabinose action. Based on the natural transformation and the inducible expression of Flp, the markerless single-gene knockout mutants of Δ
hsdR
, Δ
neuA2
, Δ
espP2
, Δ
apd
, and Δ
nanH
were constructed. In addition, a five-gene knockout mutant of Δ
hsdR
Δ
neuA2
Δ
espP2
Δ
apd
Δ
nanH
was generated by successive natural transformation with five suicide plasmids. Taken together, a markerless and multiple-gene deletion system was established for
G. parasuis
in the present study for the first time. This system is simple, efficient, and easy to manipulate for
G. parasuis
; thus, our technique will substantially aid the understanding of the etiology, pathogenesis, and genetic engineering of
G. parasuis
and other bacteria that can be naturally transformed in laboratory conditions.
Key points
• Flp recombinase excised the Kan
R
gene flanked by FRT sites in
Glaesserella parasuis.
• The regulatory expression of Flp enabled a multiple-gene knockout for
G. parasuis.
• The technique will promote the understanding of Glässer’s disease pathogens.
Graphical abstract</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-022-11994-z</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Applied Genetics and Molecular Biotechnology ; Arabinose ; Bacterial vaccines ; Biomedical and Life Sciences ; Biotechnology ; Cassettes ; Diseases ; Electroporation ; Etiology ; FLP gene ; FLP recombinase ; Gene deletion ; Gene expression ; Genetic aspects ; Genetic engineering ; Genetic transformation ; Genomes ; Gram-negative bacteria ; Health aspects ; Kanamycin ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Mutants ; Pathogenesis ; Pathogens ; Plasmids ; Product development ; Suicide ; Swine ; Transformations</subject><ispartof>Applied microbiology and biotechnology, 2022-08, Vol.106 (13-16), p.5167-5178</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022</rights><rights>COPYRIGHT 2022 Springer</rights><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c453t-5f0ca3b2ac7e9ef6acadb98ca02fd4993e49f34bc9fff7cf1f654c1182a59bdc3</citedby><cites>FETCH-LOGICAL-c453t-5f0ca3b2ac7e9ef6acadb98ca02fd4993e49f34bc9fff7cf1f654c1182a59bdc3</cites><orcidid>0000-0002-2088-9201</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-022-11994-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-022-11994-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids></links><search><creatorcontrib>Xiao, Jing</creatorcontrib><creatorcontrib>Wang, Qiaochu</creatorcontrib><creatorcontrib>Xiao, Kunxue</creatorcontrib><creatorcontrib>Zhu, Wenlong</creatorcontrib><creatorcontrib>Huang, Junhao</creatorcontrib><creatorcontrib>Cai, Xuwang</creatorcontrib><creatorcontrib>Chen, Huanchun</creatorcontrib><creatorcontrib>Xu, Xiaojuan</creatorcontrib><title>Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>Glaesserella parasuis
is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for
G. parasuis
yet. To establish a markerless knockout, deleted allelic genes with kanamycin resistance (Kan
R
) cassettes were introduced into the genome of
G. parasuis
by using natural transformation with suicide plasmids. Then, the Kan
R
cassette was excised with a thermosensitive plasmid pGF conferring a constitutive Flp expression. To realize the markerless and multiple-gene knockout, plasmid pGAF was constructed by placing the Flp gene under the control of an arabinose-inducible promoter. Firstly, pGAF was introduced into
G. parasuis
by electroporation, and the marked mutants were produced following natural transformation. Finally, the Kan
R
cassette was excised from the genome by the inducible expression of Flp upon arabinose action. Based on the natural transformation and the inducible expression of Flp, the markerless single-gene knockout mutants of Δ
hsdR
, Δ
neuA2
, Δ
espP2
, Δ
apd
, and Δ
nanH
were constructed. In addition, a five-gene knockout mutant of Δ
hsdR
Δ
neuA2
Δ
espP2
Δ
apd
Δ
nanH
was generated by successive natural transformation with five suicide plasmids. Taken together, a markerless and multiple-gene deletion system was established for
G. parasuis
in the present study for the first time. This system is simple, efficient, and easy to manipulate for
G. parasuis
; thus, our technique will substantially aid the understanding of the etiology, pathogenesis, and genetic engineering of
G. parasuis
and other bacteria that can be naturally transformed in laboratory conditions.
Key points
• Flp recombinase excised the Kan
R
gene flanked by FRT sites in
Glaesserella parasuis.
• The regulatory expression of Flp enabled a multiple-gene knockout for
G. parasuis.
• The technique will promote the understanding of Glässer’s disease pathogens.
Graphical abstract</description><subject>Applied Genetics and Molecular Biotechnology</subject><subject>Arabinose</subject><subject>Bacterial vaccines</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Cassettes</subject><subject>Diseases</subject><subject>Electroporation</subject><subject>Etiology</subject><subject>FLP gene</subject><subject>FLP recombinase</subject><subject>Gene deletion</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Genetic transformation</subject><subject>Genomes</subject><subject>Gram-negative bacteria</subject><subject>Health aspects</subject><subject>Kanamycin</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Mutants</subject><subject>Pathogenesis</subject><subject>Pathogens</subject><subject>Plasmids</subject><subject>Product development</subject><subject>Suicide</subject><subject>Swine</subject><subject>Transformations</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp9ks1rFTEUxQdR8Fn9B1wF3OhiapL5yGRZin0tFAQ_1uFO5uaRvkwyJjNgu_JPN9MRyhNxdSH8zrnnklMUbxk9Z5SKj4lS3lQl5bxkTMq6fHhW7Fhd8ZK2rH5e7CgTTSka2b0sXqV0RynjXdvuil979BhhtsGTYMgI8YjRYUoE_EDGxc12clgeMkWOPuhjWGZiPdk7yBBGdA7IBBHSYhPpIeFAspWHeYngyBzBJxPiuG1YPa_cRCLqMPbWZ_x18cKAS_jmzzwrvl99-nZ5Xd5-3t9cXtyWum6quWwM1VD1HLRAiaYFDUMvOw2Um6GWssJamqrutTTGCG2YaZtaM9ZxaGQ_6OqseL_5TjH8WDDNarRJr_E9hiUp3komuo51XUbf_YXehSX6nG6lGtY1teBP1AEcKutNyMfq1VRdCMZEy0W1Uuf_oNb0OFodPBqb308EH04EmZnx53yAJSV18_XLKcs3VseQUkSjpmjzF94rRtXaC7X1QuVeqMdeqIcsqjZRyrA_YHy67j-q30BnvPE</recordid><startdate>20220801</startdate><enddate>20220801</enddate><creator>Xiao, Jing</creator><creator>Wang, Qiaochu</creator><creator>Xiao, Kunxue</creator><creator>Zhu, Wenlong</creator><creator>Huang, Junhao</creator><creator>Cai, Xuwang</creator><creator>Chen, Huanchun</creator><creator>Xu, Xiaojuan</creator><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-2088-9201</orcidid></search><sort><creationdate>20220801</creationdate><title>Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase</title><author>Xiao, Jing ; Wang, Qiaochu ; Xiao, Kunxue ; Zhu, Wenlong ; Huang, Junhao ; Cai, Xuwang ; Chen, Huanchun ; Xu, Xiaojuan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c453t-5f0ca3b2ac7e9ef6acadb98ca02fd4993e49f34bc9fff7cf1f654c1182a59bdc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Applied Genetics and Molecular Biotechnology</topic><topic>Arabinose</topic><topic>Bacterial vaccines</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Cassettes</topic><topic>Diseases</topic><topic>Electroporation</topic><topic>Etiology</topic><topic>FLP gene</topic><topic>FLP recombinase</topic><topic>Gene deletion</topic><topic>Gene expression</topic><topic>Genetic aspects</topic><topic>Genetic engineering</topic><topic>Genetic transformation</topic><topic>Genomes</topic><topic>Gram-negative bacteria</topic><topic>Health aspects</topic><topic>Kanamycin</topic><topic>Life Sciences</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Mutants</topic><topic>Pathogenesis</topic><topic>Pathogens</topic><topic>Plasmids</topic><topic>Product development</topic><topic>Suicide</topic><topic>Swine</topic><topic>Transformations</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xiao, Jing</creatorcontrib><creatorcontrib>Wang, Qiaochu</creatorcontrib><creatorcontrib>Xiao, Kunxue</creatorcontrib><creatorcontrib>Zhu, Wenlong</creatorcontrib><creatorcontrib>Huang, Junhao</creatorcontrib><creatorcontrib>Cai, Xuwang</creatorcontrib><creatorcontrib>Chen, Huanchun</creatorcontrib><creatorcontrib>Xu, Xiaojuan</creatorcontrib><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ABI/INFORM Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Business</collection><collection>ProQuest One Business (Alumni)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xiao, Jing</au><au>Wang, Qiaochu</au><au>Xiao, Kunxue</au><au>Zhu, Wenlong</au><au>Huang, Junhao</au><au>Cai, Xuwang</au><au>Chen, Huanchun</au><au>Xu, Xiaojuan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><date>2022-08-01</date><risdate>2022</risdate><volume>106</volume><issue>13-16</issue><spage>5167</spage><epage>5178</epage><pages>5167-5178</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Glaesserella parasuis
is an important bacterial pathogen that affects the swine industry worldwide. Research on the pathogenic mechanism and genetically engineered vaccine remains undeveloped because an effective markerless and multiple-gene knockout system is unavailable for
G. parasuis
yet. To establish a markerless knockout, deleted allelic genes with kanamycin resistance (Kan
R
) cassettes were introduced into the genome of
G. parasuis
by using natural transformation with suicide plasmids. Then, the Kan
R
cassette was excised with a thermosensitive plasmid pGF conferring a constitutive Flp expression. To realize the markerless and multiple-gene knockout, plasmid pGAF was constructed by placing the Flp gene under the control of an arabinose-inducible promoter. Firstly, pGAF was introduced into
G. parasuis
by electroporation, and the marked mutants were produced following natural transformation. Finally, the Kan
R
cassette was excised from the genome by the inducible expression of Flp upon arabinose action. Based on the natural transformation and the inducible expression of Flp, the markerless single-gene knockout mutants of Δ
hsdR
, Δ
neuA2
, Δ
espP2
, Δ
apd
, and Δ
nanH
were constructed. In addition, a five-gene knockout mutant of Δ
hsdR
Δ
neuA2
Δ
espP2
Δ
apd
Δ
nanH
was generated by successive natural transformation with five suicide plasmids. Taken together, a markerless and multiple-gene deletion system was established for
G. parasuis
in the present study for the first time. This system is simple, efficient, and easy to manipulate for
G. parasuis
; thus, our technique will substantially aid the understanding of the etiology, pathogenesis, and genetic engineering of
G. parasuis
and other bacteria that can be naturally transformed in laboratory conditions.
Key points
• Flp recombinase excised the Kan
R
gene flanked by FRT sites in
Glaesserella parasuis.
• The regulatory expression of Flp enabled a multiple-gene knockout for
G. parasuis.
• The technique will promote the understanding of Glässer’s disease pathogens.
Graphical abstract</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><doi>10.1007/s00253-022-11994-z</doi><tpages>12</tpages><orcidid>https://orcid.org/0000-0002-2088-9201</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2022-08, Vol.106 (13-16), p.5167-5178 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2691788188 |
| source | SpringerLink Journals - AutoHoldings |
| subjects | Applied Genetics and Molecular Biotechnology Arabinose Bacterial vaccines Biomedical and Life Sciences Biotechnology Cassettes Diseases Electroporation Etiology FLP gene FLP recombinase Gene deletion Gene expression Genetic aspects Genetic engineering Genetic transformation Genomes Gram-negative bacteria Health aspects Kanamycin Life Sciences Microbial Genetics and Genomics Microbiology Mutants Pathogenesis Pathogens Plasmids Product development Suicide Swine Transformations |
| title | Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-08T19%3A24%3A11IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_proqu&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Generation%20of%20markerless%20and%20multiple-gene%20knockout%20in%20Glaesserella%20parasuis%20based%20on%20natural%20transformation%20and%20Flp%20recombinase&rft.jtitle=Applied%20microbiology%20and%20biotechnology&rft.au=Xiao,%20Jing&rft.date=2022-08-01&rft.volume=106&rft.issue=13-16&rft.spage=5167&rft.epage=5178&rft.pages=5167-5178&rft.issn=0175-7598&rft.eissn=1432-0614&rft_id=info:doi/10.1007/s00253-022-11994-z&rft_dat=%3Cgale_proqu%3EA711762732%3C/gale_proqu%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2695185472&rft_id=info:pmid/&rft_galeid=A711762732&rfr_iscdi=true |