酵母とEscherichia coliのゲノムを1つのサイトで分割する

M Koob¹, W Szybalski

¹McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.

Science (New York, N.Y.)

|October 12, 1990

まとめ

研究者らは,アキレスの割れ方 (AC) 方法を開発し,酵母や大腸菌などの大きなゲノムを特定の部位で正確に切断しました. この技術は,ゲノムマッピングと分子解剖のための新しいツールを提供します.

科学分野:

ゲノミクスゲノミクスとは
分子生物学は分子生物学である.
バイオテクノロジーバイオテクノロジー

背景:

大型ゲノムの精密な操作は,分子生物学にとって極めて重要です.
既存の制限酵素は,標的型ゲノム分裂の特異性が限られている.
特定のDNA分裂のための新しい方法の開発は,ゲノム分析に不可欠です.

研究の目的:

標的型ゲノム分割のためのアキレス ?? 部切断 (AC) プロセスを実証します.
マルチメガベースゲノムの精密な分子解剖のためにACを適応させる.
制限酵素の特異性を拡張するための新しいツールを作成する.

主な方法:

ラクオペレータ (lacOs) をSaccharomyces cerevisiaeとEscherichia coliのゲノムに導入する.
LACOSの部位を保護するには,LAC抑制剤 (LACI) とHAIメチルトランスフェラーゼを使用します.
保護と脱保護のステップの後,Hae IIを使用して,LacOsの部位に特別の染色体を割る.

主要な成果:

酵母とE. coliのゲノムをAC手順で,事前に決められた単一の部位で完全に切断する.
エンジニアリングされたLACOSの部位をHae II消化から保護した.
大型DNA分子の特定の割れ目のためのACの実現可能性が実証されました.

結論:

ACアプローチは,自然に存在する制限酵素の特異性を効果的に拡張します.
ACは,大型ゲノムのマッピングと精密な分子解剖のための強力な新しいツールを提供します.
この方法により,高度なゲノム研究にとって極めて重要な標的型割れが可能になります.

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