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利用fosmid载体构建的红树林功能宏基因组文库的高产量

High yield of functional metagenomic library from mangroves constructed in fosmid vector.

作者信息

Gonçalves A C S, dos Santos A C F, dos Santos T F, Pessoa T B A, Dias J C T, Rezende R P

机构信息

Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil.

Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brasil

出版信息

Genet Mol Res. 2015 Oct 2;14(4):11841-7. doi: 10.4238/2015.October.2.17.

Abstract

In the present study, metagenomic technique and fosmid vectors were used to construct a library of clones for exploring the biotechnological potential of mangrove soils by isolation of functional genes encoding hydrolytic enzymes. The library was built with genomic DNA from the soil samples of mangrove sediments and the functional screening of 1824 clones (~64 Mbp) was performed to detect the hydrolytic activity specific for cellulases, amylases (at acidic, neutral and basic pH), lipases/esterases, proteases, and nitrilases. Significant numbers of clones, positive for the tested enzyme activities were obtained. Our results indicate the importance and biotechnological potential of mangrove soils especially when compared to those obtained using other soil metagenomic libraries.

摘要

在本研究中,宏基因组技术和fosmid载体被用于构建一个克隆文库,通过分离编码水解酶的功能基因来探索红树林土壤的生物技术潜力。该文库由红树林沉积物土壤样品的基因组DNA构建而成,并对1824个克隆(约64 Mbp)进行了功能筛选,以检测对纤维素酶、淀粉酶(在酸性、中性和碱性pH条件下)、脂肪酶/酯酶、蛋白酶和腈水解酶具有特异性的水解活性。获得了大量对所测试酶活性呈阳性的克隆。我们的结果表明了红树林土壤的重要性和生物技术潜力,特别是与使用其他土壤宏基因组文库获得的结果相比。

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