Name: protein misfolding cyclic amplification of prions
Uploaded: 2012-11-07T12:00:00
Description: Watch this Scientific Journal Video about Protein Misfolding Cyclic Amplification of Prions at JoVE.com

Biz yazının eleştirel okuma için Dr Vesper Fe Marie Ramos teşekkür etmek istiyorum. Bu çalışma Araştırma Kaynakları Ulusal Merkezi (P20 RR0115635-6, C06 RR17417-01 ve G20RR024001) ve Ulusal Nörolojik Bozukluklar Enstitüsü ve İnme (2R01 NS052609) tarafından desteklenmiştir.

1. Ekipman hazırlanması <ol><li> Bir Misonix 3000 veya 37 sabit bir sıcaklıkta tutmak için bir Thermo Electron NESLAB EX-7 su banyosu (Newington, NH) bağlı Misonix 4000 sonikatör (Farmingdale, NY) ° C kullanın Thermo Scientific (Waltham, MA) dan elde edilen kubbeli kapak ile 200 ul ince duvarlı tüp PCR şeritler örnekleri sonikasyon. </li><li> Yepyeni sonikatör sürekli çalışma 9 bir &quot;soygun&quot; dönemi gerektirir. Bir iki ay aradan-in dönemi 10 ayarlanmış genlik seviyesi ile 40 s...

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+enzymatic+treatment+of+soil-bound+prions+effectively+inhibits+replication.">An enzymatic treatment of soil-bound prions effectively inhibits replication.</a> Appl. Environ. Microbiol. 77, 4313-4317 (2011).</li><li>Saunders, S. E., Shikiya, R. A., Langenfeld, K., Bartelt-Hunt, S. L., Bartz, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replication+efficiency+of+soil-bound+prions+varies+with+soil+type.">Replication efficiency of soil-bound prions varies with soil type.</a> Journal of virology. , (2011).</li><li>Schutt, C. R., Bartz, J. 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Yükseltici bulaşıcı prion proteinlerin Zorluklar in vivo deneyler uzun inkübasyon süreleri ve giderleri vardır. PMCA tekniği bulaşıcı prion ajanları güçlendirmek için uygun maliyetli bir yoludur. Birkaç laboratuar doğru vitro 7, 9, 12, 14, 19,24 içinde prion suşları yükseltmek için PMCA yeteneğini doğruladı. Prion hastalıkları türler arasında aktarılabilir. Bessen ve Marsh etkili iki ayrı hamsterden türetilmiş prion suşl...

<table border="1"><tbody><tr><td> Reaktif / Ekipman </td><td> Üretici </td><td> Kedi. Numara </td></tr><tr><td> Misonix 3000 </td><td> Misonix </td><td> S-3000 </td></tr><tr><td> Misonix 4000 </td><td> Misonix </td><td> S-4000 </td></tr><tr><td> Tenbroeck Doku Öğütücü </td><td> Kontes </td><td> 885000-0007 </td></tr><tr><td> NESLAB EX-7 Su Banyosu </td><td> Thermo Electron </td><td> NESLAB EX-7 </td></tr><tr><td> 0.2 ml PCR Tüp Şeritler </td><td> Thermo Scientific </td><td> AB-0451 </td></tr><tr><td> Triton X-100 </td><td> Sigma Aldrich </td><td> T9284-100ML </td></tr><tr><td> Komple Proteaz İnhibitör </td><td> Roche </td><td> 11 697 498 001 </td></tr><tr><td> EDTA </td><td> JT Baker </td><td> 4040-00 </td></tr><tr><td> DPBS </td><td> Mallinckrodt Baker Mediatech </td><td> 21-031-CV </td></tr><tr><td> Versi-Kuru Lab soakers </td><td> Fisher Scientific </td><td> 14 206 28 </td></tr><tr><td> Repti Therm Isıtıcı </td><td> Zoo Med Laboratories, Inc </td><td> RH-4 </td></tr></tbody></table>

protein misfolding cyclic amplification of prions

Prionlar hayvan ve insanlarda 9,18 olarak kaçınılmaz ölümcül bulaşıcı süngerimsi ensefalopati (TSE) neden bulaşıcı ajanlardır. Prion proteini iki farklı izoformu, bulaşıcı olmayan ana-kodlanmış protein (PrP C) ve enfeksiyöz protein (PrP Sc), PrP C 8 anormal katlanmış izoformu vardır. Prion ajanlar ile çalışan zorluklardan biri öncesinde ev sahibi inokülasyon 13 sonrasında klinik bulguların gelişimi uzun bir kuluçka dönemidir. Bu geleneksel olarak uzun ve pahalı hayvan biyoassay çalışmaları görevlendirdi. Ayrıca, PrP Sc biyokimyasal ve biyofiziksel özelliklere kötü onların sıradışı konformasyon ve agregasyonu devletler nedeniyle karakterize edilir. PrP Sc in vitro 14 PrP Sc PrP C tohum dönüşüm yapabilirsiniz. PMCA Adva alır bir in vitro bir tekniktirPrP C ve PrP Sc tohum 19 dakika miktarları aşan miktarda içeren bir sistem, giderek artan bir oranda, PrP Sc büyük miktarlarda üretmek için sonication ve inkübasyon döngüleri kullanarak bu yeteneği ntage. Bu teknik, etkili prion zorlanma girişime taklit etmek, PrP C PrP Sc dönüştürme türleri ve gerilme özgüllük özetlemek için, ve enfekte dokuları, sıvıları ve çevresel örneklerde 6,7,16 gelen PrP Sc çok düşük seviyelere yükseltmek için kanıtlanmıştır 23. , Kontaminasyonu en aza tutarlı sonuçlar üreten, ve bu sonuçların ölçülmesi için öneriler içeren bu kağıdı ayrıntıları PMCA protokolü. Biz de bulaşıcı prion suşları, prion zorlanma girişim, ve çevre Prionların algılama üretimi ve karakterizasyonu gibi çeşitli PMCA uygulamaları tartışmak.

Protein misfolding siklik amplifikasyon (PMCA) İn vitro 7 PrP, Sc, 12, 14, 19, 24, yükseltmek için kullanılır. Başarılı amplifikasyon PrP Sc, Şekil 3&#39;te gösterildiği gibi PK-dirençli prion proteini (hamsterden türetilmiş prion suşları için 19 ila 30 kDa geçiş) Western blot üzerinde bant yoğunluğunda bir artış ile gösterilmiştir. Kendi bant yoğunluk artışı sonra PMCA PK-dirençli PrP Sc malzeme amplifika...

Watch this Scientific Journal Video about Protein Misfolding Cyclic Amplification of Prions at JoVE.com

Protein Misfolding Cyclic Amplification of Prions

Protein misfolding siklik amplifikasyon (PMCA) Prion dönüşüm ve zorlanma ve tür engellerin çalışma için bir in vitro testtir. Aynı zamanda, bir prion saptama sistemi olarak da kullanılabilir.

Prionlar protein misfolding Döngüsel Amplifikasyon

Prions  are infectious agents that cause the inevitably fatal transmissible spongiform  encephalopathy (TSE) in animals and humans9,18. The prion protein ...

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