Name: high-throughput yeast plasmid overexpression screen
Uploaded: 2011-07-27T13:00:00
Description: Watch this Scientific Journal Video about High-throughput Yeast Plasmid Overexpression Screen at JoVE.com

이 작품은 존스 홉킨스에​​서 ALS 연구 팩커드 센터 (ADG), NIH 원장의 새로운 이노 수상 1DP2OD004417 - 01 (ADG), NIH R01 NS065317 (ADG), 리타 앨런 재단 학술 상에서 교부금에 의해 지원되었다. ADG는 퓨 자선 트러스트가 지원하는 의생명 과학 퓨 학술 있습니다.

1. 효모 변환을위한 준비 이 프로토콜은 열 96 - 웰 플레이트를 위해 설계되었습니다하지만, 이에 따라 위 또는 아래로 확장할 수 있습니다. 우리는이 프로토콜은 변화 라운드 당보다 20 96 - 웰 플레이트에 대해 제대로 작동하지 않는 것으로 확인되었습니다. 전체 변환 절차 (단계 I.3에서) 약 여덟 시간이나 걸립니다. <ol><li> 나누어지는 Biorobot의 RapidPlate 액체 처리기와 함께 96 ...

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여기 우리는 효모에서 높은 처리량 플라스미드 overexpression 화면을 수행하는 프로토콜을 제시한다. 이러한 접근 방식은 여러 세포 phenotypes의 유전자 조절에 대한 신속하고 불편 검사 수 있습니다. 이 방법을 사용하여 연구원이 주 만에 효모 게놈의 상당 부분을 스크린 수 있습니다. 이러한 불편 접근법은 또한 이전의 연구 결과에 따라 예측되지 않았을 수 있습니다 수정자의 식별을 위해 수 있습니...

<table border="1"><tbody><tr><th> 시약의 이름 </th><th> 회사 </th><th> 카탈로그 번호 </th></tr><tr><td> BioRobot RapidPlate </td><td> Qiagen </td><td> 9,000,490 </td></tr><tr><td> 96 볼트 레플리케이터 (frogger) </td><td> V &amp; P 과학 </td><td> VP404 </td></tr><tr><td> FLEXGene ORF 도서관 </td><td> Proteomics 연구소, 하버드 메디컬 스쿨 </td><td></td></tr><tr><td> 테이블탑 원심 분리기 </td><td> Eppendorf </td><td> 5810R </td></tr><tr><td> 500mL 플라스크 실패하는 </td><td> Bellco </td><td> 2543-00500 </td></tr><tr><td> 2.8L 트리플 당황하게 Fernbach 플라스크 </td><td> Bellco </td><td> 2551-02800 </td></tr><tr><td> 100μL Rapidplate 피펫 팁 </td><td> Axygen </td><td> ZT - 100 - RS </td></tr><tr><td> 200μL Rapidplate 피펫 팁 </td><td> Axygen </td><td> ZT - 200 - RS </td></tr></tbody></table>

high-throughput yeast plasmid overexpression screen

신진 효모, Saccharomyces cerevisiae의는 인간의 질병에 직접 관련성이있는 사람 등 많은 중요한 세포 과정의 기본 메커니즘을 정의하는 강력한 모델 시스템입니다. 워낙 짧은 세대 시간과 잘 특징 게놈의 효모 모델 시스템의 주요 실험 이점은 주어진 과정에 관련된 유전자 및 경로를 식별하는 유전자 화면을 수행할 수있는 능력입니다. 지난 30 년 동안 같은 유전자 화면은 세포주기, 분비 경로와 진핵 세포 생물학 1-5의 많은 높은 보존 측면을 명료하게하다하는 데 사용되었습니다. 지난 몇 년 동안, 효모 종자와 plasmids 여러 genomewide 라이브러리는 60-10을 생성되었습니다. 이 컬렉션은 이제 이득과 손실 함수 접근법 11-16을 사용하여 유전자 기능의 체계적인 심문을 위해 수 있습니다. 여기 우리는 5500 효모 plasmids의 못하였 느니라 라이브러리를 사용하여 플라스미드 overexpression 화면을 수행하는 액체 처리 로봇과 높은 처리량 효모 변환 프로토콜의 사용에 대한 자세한 프로토콜을 제공합니다. 우리는 집합을 일으키는 인간 neurodegenerative 질병 단백질의 축적과 관련된 독성의 유전자 조절을 식별이 화면을 사용하고 있습니다. 여기에 제시 방법은 관심의 다른 세포 phenotypes의 연구에 쉽게 적용할 수 있습니다.

Watch this Scientific Journal Video about High-throughput Yeast Plasmid Overexpression Screen at JoVE.com

High-throughput Yeast Plasmid Overexpression Screen

여기에 플라스미드 overexpression 화면 설명<em&gt; Saccharomyces cerevisiae</em&gt; 액체 처리 로봇과 못하였 느니라 플라스미드 라이브러리 및 높은 처리량 효모 변환 프로토콜을 사용합니다.

높은 처리량 효모 플라스미드 Overexpression 화면

The budding yeast, Saccharomyces cerevisiae, is a powerful model system for defining fundamental mechanisms of many important cellular processes, ...

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