Name: rapid deletion production in fungi via agrobacterium mediated transformation of oscar deletion constructs
Uploaded: 2017-06-12T13:00:00
Description: Watch this Scientific Journal Video about Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs at JoVE.com

Forfatterne takker følgende grundskole- og gymnasieelever for deres arbejde med at generere OSCAR-mutanter i Fusarium verticillioiodes : Anjellica Miller, Athar Naseer, Xiu Lin, Katelyn Woodburry, Chelsea Patterson, Kathleen Robertson, Krystina Bradley, Ashton Rogers, Alexis McKensie, Manny Hernandez , Ashli ​​Crepsac, Jeff Delong, Christian King, Gi Jeong, Maria Belding, Christy Burre, Daniel O&#39;Meara, Lauren (Victoria) Cook, Jake Goodman, Sampriti De, Oge Okoye, Alyssa Beckstead, Garrett Hibbs, Nick Goldstein, Caroline Twum , Chris Benson, Louis Stokes, Hannah Itell, Jane Hulse, Jasim Mohammed, James Loggins, Kelli Russell, Gre&#39;Nisha Jones, Kristin Sheaffer, Mariam Hammady, Ava Wilson, Katrina Bazemore, Toney Harper, Karlin McGhee, Mohmed Momin, Rima Momin , Thi Ngoc Le og Angel Pham.

1. Primer Design for PCR Amplification of Gene Flanks <ol><li> Download til en tekstbehandlingsfil den genomiske region af genet af interesse (GOI), herunder den åbne læseramme (ORF) og mindst 2 kb flankerer genet på hver side fra FungiDB eller anden genomisk datafilde. </li><li> Fremhæv ORF&#39;en beregnet til sletning og etiket start og stop kodoner. </li><li> Identificer og markér tilstødende ORF&#39;er inden for den downloadede sekvens. </li><li> Brug 2-kb 5&#39;-enden af ​​GOI ORF og primerdesig...

<ol>
	<li>Paz, Z., Garc&#237;a-Pedrajas, M. D., Andrews, D. L., Klosterman, S. J., Baeza-Monta&#241;ez, L., Gold, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One+step+construction+of+Agrobacterium-Recombination-ready-plasmids+(OSCAR),+an+efficient+and+robust+tool+for+ATMT+based+gene+deletion+construction+in+fungi.">One step construction of Agrobacterium-Recombination-ready-plasmids (OSCAR), an efficient and robust tool for ATMT based gene deletion construction in fungi.</a> Fungal Genet Biol. 48 (7), 677-684 (2011).</li><li>Gibson, D. G., Young, L., Chuang, R. Y., Venter, J. C., Hutchison, C. A., Smith, H. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enzymatic+assembly+of+DNA+molecules+up+to+several+hundred+kilobases.">Enzymatic assembly of DNA molecules up to several hundred kilobases.</a> Nat Methods. 6 (5), 343-345 (2009).</li><li>Klosterman, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+genomics+yields+insights+into+niche+adaptation+of+plant+vascular+wilt+pathogens.">Comparative genomics yields insights into niche adaptation of plant vascular wilt pathogens.</a> PLoS Pathog. 7 (7), (2011).</li><li>Xue, C., Wu, D., Condon, B. J., Bi, Q., Wang, W., Turgeon, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+gene+knockout+in+the+maize+pathogen+Setosphaeria+turcica+using+Agrobacterium+tumefaciens-mediated+transformation.">Efficient gene knockout in the maize pathogen Setosphaeria turcica using Agrobacterium tumefaciens-mediated transformation.</a> Phytopathology. 103 (6), 641-647 (2013).</li><li>Xu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-throughput+gene+disruption+methodology+for+the+entomopathogenic+fungus+Metarhizium+robertsii.">A high-throughput gene disruption methodology for the entomopathogenic fungus Metarhizium robertsii.</a> PloS One. 9 (9), (2014).</li><li>Crutcher, F. K., Liu, J., Puckhaber, L. S., Stipanovic, R. D., Bell, A. A., Nichols, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FUBT,+a+putative+MFS+transporter,+promotes+secretion+of+fusaric+acid+in+the+cotton+pathogen+Fusarium+oxysporum+f.+sp.+vasinfectum.">FUBT, a putative MFS transporter, promotes secretion of fusaric acid in the cotton pathogen Fusarium oxysporum f. sp. vasinfectum.</a> Microbiology. 161, 875-883 (2015).</li><li>Yu, X., Wang, Y., Pan, J., Wei, D., Zhu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+frequency+of+homologous+gene+disruption+by+single-stranded+DNA+in+the+taxol-producing+fungus+Pestalotiopsis+microspora.">High frequency of homologous gene disruption by single-stranded DNA in the taxol-producing fungus Pestalotiopsis microspora.</a> Ann Microbiol. 65 (4), 2151-2160 (2015).</li><li>Korn, M., Schmidpeter, J., Dahl, M., M&#252;ller, S., Voll, L. M., Koch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Genetic+Screen+for+Pathogenicity+Genes+in+the+Hemibiotrophic+Fungus+Colletotrichum+higginsianum+Identifies+the+Plasma+Membrane+Proton+Pump+Pma2+Required+for+Host+Penetration.">A Genetic Screen for Pathogenicity Genes in the Hemibiotrophic Fungus Colletotrichum higginsianum Identifies the Plasma Membrane Proton Pump Pma2 Required for Host Penetration.</a> PloS One. 10 (5), e0125960 (2015).</li><li>Chettri, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Regulation of dothistromin toxin biosynthesis by the pine needle pathogen Dothistroma septosporum: a thesis presented in the partial fulfilment of the requirements for the degree of Doctor of Philosophy (PhD) in Genetics at Massey University, Manawatu, New Zealand . , (2014).</li><li>Chen Zhou, ., Yujun Yang, ., Jong, A. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mini-prep+in+ten+minutes.">Mini-prep in ten minutes.</a> Biotechniques. 8 (2), 172 (1990).</li><li>Sanger, F., Nicklen, S., Coulson, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+sequencing+with+chain-terminating+inhibitors.">DNA sequencing with chain-terminating inhibitors.</a> P Natl Acad SciUSA. 74 (12), 5463-5467 (1977).</li><li>Khang, C. H., Park, S. Y., Rho, H. S., Lee, Y. H., Kang, S., Wang, K. a. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Filamentous+fungi+(Magnaporthe+grisea+and+Fusarium+oxysporum).">Filamentous fungi (Magnaporthe grisea and Fusarium oxysporum).</a> Agrobacterium Protocols. 2, 403-420 (2007).</li><li>Zhang, Y. J., Zhang, S., Liu, X. Z., Wang Wen, H. A., M, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+of+genomic+DNA+extraction+suitable+for+analysis+of+bulk+fungal+strains.">A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains.</a> Lett Appl Microbiol. 51 (1), 114-118 (2010).</li><li>Pluthero, F. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+purification+of+high-activity+Taq+DNA+polymerase.">Rapid purification of high-activity Taq DNA polymerase.</a> Nucleic Acids Res. 21 (20), 4850-4851 (1993).</li><li>McCluskey, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Boosting+Research+and+Industry+by+Providing+Extensive+Resources+for+Fungal+Research.">Boosting Research and Industry by Providing Extensive Resources for Fungal Research.</a> Gene Expression Systems in Fungi: Advancements and Applications. , 361-384 (2016).</li></ol>

One-Step Konstruktion af Agrobacterium- Recombination-ready-plasmids (OSCAR) har med succes været anvendt med et stadigt stigende antal Ascomycota svampe. Metoden bør også let kunne anvendes på Basidiomycota og arter fra andre svampepyla (med passende promotorer, der kører selekterbare markørgener), forudsat at Agrobacterium- medieret transformation og homolog rekombination er mulige. Yderligere markørvektorer er blevet dannet for at diversificere valg af anti-svampeforbindelse samt tillade frem...

Genetisk dissektion er en kraftfuld metode til bestemmelse af den funktionelle betydning af individet eller kombinationer af gener. En standard tilgang til at forstå rollen af ​​specifikke gener er produktion af enkeltgenmutanter uændret i ethvert andet gen. Den mest kraftfulde og mindst potentielt konfronterende tilgang er fuldstændig og præcis deletion af et gen af ​​interesse er åben læseramme (GOI ORF) uden skade på nogen anden genfunktion. Fordi standardiseringsmetoder til sletning af plasmidgenerering kræver flere trin, var det rationelle for OSCAR 1 at producere en hurtigere in vitro- tilgang. Figur 1 viser samleprocessen i OSCAR-tilgangen. Fremgangsmåden beskrevet her har den fordel at kombinere hurtig konstruktion af individuelle gendeletionsvektorer i en enkelt multipartreaktion i kombination med efterfølgende Agrobacterium tumefaciens- medieret transfoRmation (ATMT). OSCAR er meget hurtig og sammenligner sig godt med andre strategier, såsom anvendelse af Gibson-samling i gær 2 . OSCAR-metoden er blevet anvendt succesfuldt med flere Ascomycota-svamparter. Disse arter omfatter: Fusarium verticillioides (upubliceret), Verticillium dahliae 3 , Setosphaeria turcica 4 , Metarhizium robertsii 5 , Fusarium oxysporum f. sp. Vasinfectum 6 , Pestalotiopsis microspora 7 , Colletotrichum higginsianum 8 og Dothistroma septosporum 9 og Sarocladium zeae (upubliseret) . Denne protokol tilvejebringer trin-for-trin instruktion til fremgangsmåden inklusiv primer design, flank PCR amplifikation, OSCAR BP reaktion, deletion konstruktion struktur bekræftelse, transformatIon af Agrobacterium med konstruktionen efterfulgt af ATMT-baseret overførsel af deletionskonstruktionen i svampecellerne og endelig differentiering af svampe deletionsmutanter fra dem med ektopisk integrerede deletionskonstruktioner.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>FungiDB</td>
			<td></td>
			<td></td>
			<td>Database/ http://fungidb.org/fungidb/</td>
		</tr>
		<tr>
			<td>IDT PrimerQuest</td>
			<td>IDT</td>
			<td></td>
			<td>Primer design online software/ http://www.idtdna.com/Primerquest/Home/Index</td>
		</tr>
		<tr>
			<td>Microsoft Word</td>
			<td></td>
			<td></td>
			<td>Sequence file manipulation</td>
		</tr>
		<tr>
			<td>Low Na LB Spec 100 medium</td>
			<td></td>
			<td></td>
			<td>E. coli transformant selection, composition: 1% tryptone, 0.05% NaCl, 0.5% yeast extract, 1.5 % agar if for solid medium</td>
		</tr>
		<tr>
			<td>Co-cultivation medium</td>
			<td></td>
			<td></td>
			<td>ATMT transformation induction (Reference 12)</td>
		</tr>
		<tr>
			<td>Aspergillus minimal medium with Hygromycin</td>
			<td></td>
			<td></td>
			<td>Fungal transformant selection</td>
		</tr>
		<tr>
			<td>PDA medium</td>
			<td>Acumedia</td>
			<td>7149A</td>
			<td>Single spore slant tubes</td>
		</tr>
		<tr>
			<td>PDA-Hyg-Kan medium</td>
			<td></td>
			<td></td>
			<td>Fungal ransformant isolation, PDA containing 150 &#956;g/ml hygromycin B and 100 &#956;g/ml Kanamycin;</td>
		</tr>
		<tr>
			<td>Glass beads</td>
			<td>Genlantis</td>
			<td>C400100</td>
			<td>Plate spreading</td>
		</tr>
		<tr>
			<td>Nitrocellulose filters (47mm)</td>
			<td>Fisher</td>
			<td>09-719-555</td>
			<td>Co-culturing for ATMT</td>
		</tr>
		<tr>
			<td>Various centifuge tubes</td>
			<td></td>
			<td></td>
			<td>multiple preps</td>
		</tr>
		<tr>
			<td>Petri plates (various)</td>
			<td></td>
			<td></td>
			<td>Culturing of bacteria and Fungi</td>
		</tr>
		<tr>
			<td>pA-Hyg OSCAR</td>
			<td>Addgene</td>
			<td>29640</td>
			<td>Selectable marker vector</td>
		</tr>
		<tr>
			<td>pOSCAR</td>
			<td>Addgene</td>
			<td>29639</td>
			<td>Assembly vector</td>
		</tr>
		<tr>
			<td>DH5a One Shot Competent E. coli cells</td>
			<td>Life Technologies&#160;</td>
			<td>12297-016</td>
			<td>BP reaction transformation</td>
		</tr>
		<tr>
			<td>ccdB survival E. coli cells</td>
			<td>Life Technologies&#160;</td>
			<td>A10460</td>
			<td>Maintenance of pOSCAR</td>
		</tr>
		<tr>
			<td>Wooden transfer sticks</td>
			<td></td>
			<td></td>
			<td>Colony streaking</td>
		</tr>
		<tr>
			<td>Toothpicks</td>
			<td></td>
			<td></td>
			<td>Colony picking</td>
		</tr>
		<tr>
			<td>Microcentrifuge</td>
			<td></td>
			<td></td>
			<td>Pelleting Bacteria etc</td>
		</tr>
		<tr>
			<td>Preparative centrifuge</td>
			<td></td>
			<td></td>
			<td>Fungal spore collection</td>
		</tr>
		<tr>
			<td>Dissecting microscope</td>
			<td></td>
			<td></td>
			<td>Single spore isolation</td>
		</tr>
		<tr>
			<td>Automated Cell Counter</td>
			<td></td>
			<td></td>
			<td>Spore suspension calculation</td>
		</tr>
		<tr>
			<td>Compound microscope</td>
			<td></td>
			<td></td>
			<td>Hemocytometer cell counting</td>
		</tr>
		<tr>
			<td>QIAquick PCR Purification Kit&#160;</td>
			<td>Qiagen</td>
			<td>28104</td>
			<td>PCR gene flank produict purification</td>
		</tr>
		<tr>
			<td>TaKaRa LA Taq&#160;</td>
			<td>Takara Bio USA</td>
			<td>RR002A</td>
			<td>Hi Fidelity taq polymerase for OSCAR flank generation</td>
		</tr>
		<tr>
			<td>Hygromycin B</td>
			<td>InvivoGen</td>
			<td>ant-hg-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectinomycin</td>
			<td>Sigma</td>
			<td>22189-32-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Cefotaxim&#160;</td>
			<td>TCI America</td>
			<td>C2224</td>
			<td></td>
		</tr>
		<tr>
			<td>Moxalactam&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>43963</td>
			<td></td>
		</tr>
		<tr>
			<td>GelRed</td>
			<td>&#160;Phenix Research Products</td>
			<td>RGB-4103</td>
			<td>Post staining agarose gels</td>
		</tr>
		<tr>
			<td>Qiagen QIAquick PCR Purification Kit (Cat. No. 28104)&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>(OneShot_ Mach1TM T1R or One Shot_ OmniMAX&#8482; 2 T1R from Invitrogen)&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>C862003</td>
			<td></td>
		</tr>
		<tr>
			<td>Gateway BP Clonase II Enzyme mix</td>
			<td>Thermo Fisher Scientific</td>
			<td>11789020</td>
			<td>Used to assemble deletion construct in pOSAR</td>
		</tr>
		<tr>
			<td>PrimerQuest tool</td>
			<td>IDT&#160;</td>
			<td></td>
			<td>Used in step 1.4; available on http://www.idtdna.com/Primerquest/Home/Index</td>
		</tr>
	</tbody>
</table>

rapid deletion production in fungi via agrobacterium mediated transformation of oscar deletion constructs

Præcis sletning af gen (er) af interesse, medens resten af ​​genomet uændret giver det ideelle produkt til at bestemme det pågældende gens funktion i den levende organisme. I denne protokol beskrives OSCAR-metoden for præcis og hurtig deletionsplasmidkonstruktion. OSCAR er afhængig af kloningsystemet, hvori en enkelt rekombinase-reaktion udføres, der indeholder de oprensede PCR-amplificerede 5&#39;- og 3&#39;-flanker af genet af interesse og to plasmider, pA-Hyg OSCAR (markørvektoren) og pOSCAR vektor). Bekræftelse af den korrekt monterede deletionsvektor udføres ved hjælp af restriktionsfordøjelsesmapping efterfulgt af sekventering. Agrobacterium tumefaciens anvendes derefter til at mediere introduktion af deletionskonstruktionen i svampesporer (benævnt ATMT). Endelig beskrives et PCR-assay for at bestemme, om deletionskonstruktionen integreres ved homolog eller ikke-homolog rekombination, indikerende gendeletion ellerEktopisk integration, henholdsvis. Denne fremgangsmåde er blevet anvendt til sletning af talrige gener i Verticillium dahliae og i Fusarium verticillioides blandt andre arter.

OSCAR-metoden genererer i en enkelt reaktion et plasmid indeholdende flankerne af målgenet, der skal slettes omkring den selekterbare markørkassette. Produktionen af ​​sletningskonstruktioner under anvendelse af OSCAR er meget effektiv. Systemet kan imidlertid producere partielle konstruktioner indeholdende nogle, men ikke alle tre fragmenter (de to genflanker og den selekterbare markør). Generelt indeholder størstedelen af E. coli transformanter den korrekte OSCAR konst...

Watch this Scientific Journal Video about Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs at JoVE.com

Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs

Gendeletionsmutanter frembragt ved homolog rekombination er guldstandarden for genfunktionsstudier. OSCAR (One Step Construction of Agrobacterium-Recombination-ready-plasmids) fremgangsmåden til hurtig generering af deletionskonstruktioner er beskrevet. Agrobacterium- medieret svamptransformation følger. Endelig præsenteres en PCR-baseret bekræftelsesmetode for gendeletioner i svamptransformanter.

Hurtig sletningsproduktion i svampe via<em&gt; Agrobacterium</em&gt; Medieret transformation af OSCAR-sletningskonstruktioner

Precise deletion of gene(s) of interest, while leaving the rest of the genome unchanged, provides the ideal product to determine that particular ...

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Rapid Deletion Production in Fungi via Agrobacterium Mediated Transformation of OSCAR Deletion Constructs

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