Substrate Induced Gene Expression Screening of Enviromental Metagenome Libraries for Isolation of Catabolic Genes  

Oleh:

Uchiyama, Taku ; Abe, Takashi ; Ikemura, Toshimichi ; Watanabe, Kazuya

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 1 (Jan. 2005), page 88-93.
Topik: genesis; gene expression; metagenome; catabolic genes

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.2 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

Recent awareness that most microorganisms in the environment are resistant to cultivation has prompted scientists to directly clone useful genes from environmental metagenomes. Two screening methods are currently available for the metagenome approach, namely, nucleotide sequence based screening and enzyme activity based screening. Here we have introduced and optimized a third option for the isolation of novel catabolic operations, that is, substrate induced gene expression screening (SIGEX). This method is based on the knowledge that catabolic gene expression is generally induced by relevant substrates and in many cases, controlled by regulatory elements situated proximate to catabolic genes. For SIGEX to be high througput, we constructed an operon trap gap expression vector available for shortgun clonung that allows for the selection of positive clones in liquid cultures by fluorescence activated cell sorting. The utility of SIGEX was demonstrated by the cloning of aromatic hydrocarbon induced genes from a groundwater metagenome library and subsequent genome informatics analysis.

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