Prospecting for novel lipase genes using PCR
Prospecting for novel lipase genes using PCR
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DOI：
10.1099/00221287-148-8-2283
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发表时间：
2002-08-01
期刊：
影响因子：
2.8
通讯作者：
Bergquist, PL
中科院分区：
文献类型：
--
作者：
Bell, PJL;Sunna, A;Bergquist, PL
介绍了一种适用于直接从环境DNA中分离脂肪酶基因的聚合酶链式反应方法。通过对保守区域的广泛分析和仔细的引物设计，与脂肪酶基因之间的低水平相似性相关的问题被克服了。利用这种方法，直接从环境DNA中分离到一个脂肪酶基因(oli-lipase)。该脂肪酶在氨基酸水平上与已知的其他脂肪酶的相似性不到20%。研究还发现，α/β水解酶超家族的远亲成员与脂肪酶有相似的保守基序，从而使这些基因成为聚合酶链反应寻找基因的靶点。
A PCR method suitable for the isolation of lipase genes directly from environmental DNA is described. The problems associated with the low levels of similarity between lipase genes were overcome by extensive analysis of conserved regions and careful primer design. Using this method, a lipase gene (oli-lipase) was isolated directly from environmental DNA. This lipase showed less than 20% similarity with other known lipases at the amino acid level. The study also revealed that distantly related members of the alpha/beta hydrolase superfamily share similar conserved motifs with the lipases, thus making these genes targets for gene prospecting by PCR.