Name: collection, isolation and enrichment of naturally occurring magnetotactic bacteria from the environment
Uploaded: 2012-11-15T11:55:00
Description: Watch this Scientific Journal Video about Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment at JoVE.com

Dette arbejde blev støttet af tilskud fra den amerikanske National Science Foundation (EAR-0.920.299 og EAR-0.745.808), US National Science Foundation East Asien og Stillehavsområdet Summer Institutes, Geologisk Society of America Research Grant Program og Alumni Tilskud til Graduate Research og Scholarship fra The Ohio State University. Vi vil gerne takke redaktøren og to anonyme korrekturlæsere for deres indsigtsfulde kommentarer.

1. MTB Collection <ol><li> Når der træffes beslutning om en ferskvands websted til at indsamle magnetotactic bakterier (MTB), er det ofte bedst at starte med en dam eller langsom strøm, der har en blød mudret sedimentlag. I denne demonstration har vi samlet en prøve på kanten af ​​Olentangy River på campus af The Ohio State University (OSU) i Columbus, Ohio (USA). Mens dette var en bekvem placering til vores demonstration, protokollen beskrevet her er gældende for alle akvatiske sted. Materialer i denne pr...

<ol>
	<li>Blakemore, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetotactic+bacteria.">Magnetotactic bacteria.</a> Science. 190, 377-379 (1975).</li><li>Blakemore, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetotactic+bacteria.">Magnetotactic bacteria.</a> Annual Reviews in Microbiology. 36, 217-238 (1982).</li><li>Bazylinski, D. A., Frankel, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+Biomineralization+of+Magnetite+(Fe3O4)+and+Greigite+(Fe3S4)+in+a+Magnetotactic+Bacterium.">Controlled Biomineralization of Magnetite (Fe3O4) and Greigite (Fe3S4) in a Magnetotactic Bacterium.</a> Applied and Environmental Microbiology. 61, 3232-3239 (1995).</li><li>Lefevre, C. T., Menguy, N., 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+Cultured+Greigite-Producing+Magnetotactic+Bacterium+in+a+Novel+Group+of+Sulfate-Reducing+Bacteria.">A Cultured Greigite-Producing Magnetotactic Bacterium in a Novel Group of Sulfate-Reducing Bacteria.</a> Science. 334, 1720-1723 (2011).</li><li>Simmons, S. L., Bazylinski, D. A., 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=South-seeking+magnetotactic+bacteria+in+the+Northern+Hemisphere.">South-seeking magnetotactic bacteria in the Northern Hemisphere.</a> Science. 311, 371-374 (2006).</li><li>Wolfe, R., Thauer, R., 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+'capillary+racetrack'+method+for+isolation+of+magnetotactic+bacteria.">A 'capillary racetrack' method for isolation of magnetotactic bacteria.</a> FEMS Microbiology Letters. 45, 31-35 (1987).</li><li>Rodgers, F. G., Blakemore, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intercellular+structure+in+a+many-celled+magnetotactic+prokaryote.">Intercellular structure in a many-celled magnetotactic prokaryote.</a> Archives of Microbiology. 154, 18-22 (1990).</li><li>Moench, T. T., Konetzka, W., 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+novel+method+for+the+isolation+and+study+of+a+magnetotactic+bacterium.">A novel method for the isolation and study of a magnetotactic bacterium.</a> Archives of Microbiology. 119, 203-212 (1978).</li><li>Balkwill, D., Maratea, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrastructure+of+a+magnetotactic+spirillum.">Ultrastructure of a magnetotactic spirillum.</a> Journal of Bacteriology. 141, 1399-1408 (1980).</li><li>Lins, U., Freitas, F., 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=Simple+homemade+apparatus+for+harvesting+uncultured+magnetotactic+microorganisms.">Simple homemade apparatus for harvesting uncultured magnetotactic microorganisms.</a> Brazilian Journal of Microbiology. 34, 111-116 (2003).</li><li>Jogler, C., Lin, W., 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=Toward+Cloning+of+the+Magnetotactic+Metagenome:+Identification+of+Magnetosome+Island+Gene+Clusters+in+Uncultivated+Magnetotactic+Bacteria+from+Different+Aquatic+Sediments.">Toward Cloning of the Magnetotactic Metagenome: Identification of Magnetosome Island Gene Clusters in Uncultivated Magnetotactic Bacteria from Different Aquatic Sediments.</a> Applied and Environmental Microbiology. 75, 3972-3979 (2009).</li><li>Lin, W., Li, 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=Newly+Isolated+but+Uncultivated+Magnetotactic+Bacterium+of+the+Phylum+Nitrospirae+from+Beijing,+China.">Newly Isolated but Uncultivated Magnetotactic Bacterium of the Phylum Nitrospirae from Beijing, China.</a> Applied and Environmental Microbiology. 78, 668-675 (2012).</li><li>Li, J., Pan, Y., 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=Biomineralization,+crystallography+and+magnetic+properties+of+bullet-shaped+magnetite+magnetosomes+in+giant+rod+magnetotactic+bacteria.">Biomineralization, crystallography and magnetic properties of bullet-shaped magnetite magnetosomes in giant rod magnetotactic bacteria.</a> Earth and Planetary Science Letters. 293, 368-376 (2010).</li><li>Oestreicher, Z., Valerde-Tercedor, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetosomes+and+magnetite+crystals+produced+by+magnetotactic+bacteria+as+resolved+by+atomic+force+microscopy+and+transmission+electron+microscopy.">Magnetosomes and magnetite crystals produced by magnetotactic bacteria as resolved by atomic force microscopy and transmission electron microscopy.</a> Micron. 43, 1331-1335 (2012).</li></ol>

Magnetotactic bakterier ikke nødvendigvis findes i alle vandmiljøet 8, men når de finder sted, kan de findes i størrelsesordenen 100 - 1000 celler per milliliter 2. For at observere MTB ved optisk mikroskopi, skal du cirka 50 bakterier / ml i din prøve 8. Hvis der ikke er eller kun få MTB i din prøve, så vil du enten nødt til at vælge en ny miljømæssig websted for at hente dine prøver eller du bliver nødt til at prøve et eller flere af de teknikker diskuteres i næste afsn...

<table border="1"><tbody><tr><td> Varenavn </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer (valgfrit) </td></tr><tr><td> Objektglas </td><td> Fisher Scientific </td><td> S95933 </td><td></td></tr><tr><td> Glas Pasteur pipetter </td><td> Fisher Scientific </td><td> 13-678-6A </td><td></td></tr><tr><td> O-ring </td><td> Hardware butik </td><td></td><td></td></tr><tr><td> Dækglas </td><td> Fisher Scientific </td><td> 12-542B </td><td></td></tr><tr><td> Stangmagnet </td><td> Fisher Scientific </td><td> S95957 </td><td></td></tr><tr><td> Container </td><td> Enhver </td><td></td><td> Enhver plastic eller glasbeholder, der kan indeholde mindst 0,5 L og kan forsegles </td></tr><tr><td> Bomuld </td><td> Enhver </td><td></td><td></td></tr><tr><td> Mikroskop med 60X tør linse </td><td> Zeiss </td><td></td><td> En 60X tør linse er ikke absolut nødvendigt, men det giver en høj NA uden brug af olie </td></tr><tr><td> Diamond pen </td><td> Fisher Scientific </td><td> 08-675 </td><td></td></tr><tr><td> 0,22 mm filter </td><td> Fisher Scientific </td><td> 09-719C </td><td></td></tr><tr><td> 1 ml sprøjte </td><td> Fisher Scientific </td><td> NC9788564 </td><td></td></tr><tr><td> Mikrocentrifugerør </td><td> Fisher Scientific </td><td> 02-681-320 </td><td></td></tr><tr><td> Formvar / Carbon200 mesh, kobber gitre </td><td> Ted Pella Inc. </td><td> 01.800 </td><td></td></tr><tr><td> Uranylacetat </td><td> Ted Pella Inc. </td><td> 19.481 </td><td></td></tr><tr><td> Tecnai Spirit TEM </td><td> FEI </td><td></td><td></td></tr><tr><td> Tecnai F20 S / TEM </td><td> FEI </td><td></td><td></td></tr></tbody></table>

collection, isolation and enrichment of naturally occurring magnetotactic bacteria from the environment

Magnetotactic bakterier (MTB) er akvatiske mikroorganismer, der først blev især beskrevet i 1975 1 fra sedimentprøver indsamlet på strandengene i Massachusetts (USA). Siden da MTB er blevet opdaget i lagdelt vand-og sediment-kolonner fra hele verden 2. Ét fælles træk for alle MTB er, at de indeholder magnetosomes, som er intracellulære, membranbundne magnetiske nanokrystaller af magnetit (Fe 3 O 4) og / eller greigite (Fe 3 S 4) eller begge dele 3, 4. På den nordlige halvkugle, er MTB typisk tiltrukket af den sydlige ende af en stangmagnet, mens der i den sydlige halvkugle, de er som regel tiltrukket af den nordlige ende af en magnet 3,5. Denne egenskab kan udnyttes, når de forsøger at isolere MTB fra miljøprøver. En af de mest almindelige måder at berige MTB er at anvende en klar plast beholder til opsamling af sediment og vand fra en naturlig kilde,såsom en ferskvand dam. På den nordlige halvkugle, er den sydlige del af en stangmagnet anbragt mod ydersiden af ​​beholderen lige over sedimentet i sediment-vand-grænsefladen. Efter nogen tid kan bakterierne fjernes fra indersiden af beholderen nær magneten med en pipette og derpå beriget yderligere ved anvendelse af en kapillær racerbane 6 og en magnet. Når beriget, bakterierne kan placeres på et objektglas under anvendelse af en hængende dråbe fremgangsmåde og observeret i et lysmikroskop eller afsættes på et kobbernet og observeret under anvendelse transmissionselektronmikroskopi (TEM). Under anvendelse af denne fremgangsmåde kan isoleret MTB undersøges mikroskopisk for at bestemme egenskaber såsom svømning adfærd, arten og antallet af flageller, cellemorfologi af cellerne, formen af ​​de magnetiske krystaller, antal magnetosomes, antal magnetosome kæder i hver celle, sammensætning af De nanomineral krystaller, og tilstedeværelse af intracellulære vakuoler.

En magnet er et effektivt værktøj, som kan anvendes til at isolere magnetotactic bakterier (MTB) indeholdt i miljøprøver (figur 1A). En kapillær racerbane (figur 1 B) anvender de magnetiske egenskaber af MTB at tiltrække dem gennem en bomuldsprop, hvor de kan adskilles fra ikke-magnetotactic mikroorganismer også indeholdt i miljøprøven. <img alt="Figur 1" src="/files/ftp_upload/50123/50123fig1.jpg" /> <stro...

Watch this Scientific Journal Video about Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment at JoVE.com

Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment

Vi viser en fremgangsmåde til at indsamle magnetotactic bakterier (MTB), der kan anvendes til naturlige vande. MTB kan isoleres og beriget fra sedimentprøver under anvendelse af en relativt enkel opsætning, der udnytter bakteriernes naturlige magnetisme. Isoleret MTB kan derefter blive undersøgt nærmere ved hjælp af både lys-og elektronmikroskopi.

Indsamling, Isolering og berigelse af naturligt forekommende Magnetotactic Bakterier fra Miljø

Magnetotactic bacteria (MTB) are aquatic microorganisms that were first notably described in 19751 from sediment samples collected in salt marshes of ...

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