Weizmann
Institute of
Science的研究人員建立了大腸桿菌菌株庫,能夠在活細胞中實時檢測基因表達。每個報告菌株都標記了明亮的、快速折疊的綠色熒光蛋白(GFP),融合到低拷貝質粒中大腸桿菌啟動子的全長拷貝上。這個文庫包括大腸桿菌K12菌株MG1655的1900多個啟動子(整個基因組有2500個啟動子),并且能夠以高精度和重復性地實時檢測基因表達。使用FACS或延時熒光顯微鏡在多孔板中進行實驗,測量單個細胞中的基因表達,靈敏度高。
1. K.A. Datsenko, B.L. Wanner, One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. PNAS. 97, 6640-6645 (2000).
2. S. Kalir et al., Ordering genes in a flagella pathway by analysis of expression kinetics from living bacteria. Science.292, 2080–2083 (2001).
3. A. Zaslaver et al., Just-in-time transcription program in metabolic pathways. Nature Genetics.36, 486–491 (2004).
4. G. Butland et al.,Interaction network containing conserved and essential protein complexes in Escherichia coli. Nature. 433, 531–537 (3 February 2005).