Retrieval of glycoside hydrolase family 9 cellulase genes from environmental DNA by metagenomic gene specific multi-primer PCR
Retrieval of glycoside hydrolase family 9 cellulase genes from environmental DNA by metagenomic gene specific multi-primer PCR
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通过宏基因组基因特异性多引物 PCR 从环境 DNA 中检索糖苷水解酶家族 9 纤维素酶基因
DOI：
10.1007/s10529-012-0855-1
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发表时间：
2012-01
影响因子：
2.7
通讯作者：
Wang, Qiuyan
中科院分区：
文献类型：
--
作者：
Xiong, Xiaolong;Yin, Xiaopu;Pei, Xiaolin;Jin, Peng;Zhang, Ao;Li, Yan;Gong, Weibo;Wang, Qiuyan
提出了一种新的方法，称为元基因组基因特异性多引物聚合酶链式反应(MGSM-PCR)，该方法使用从构建的元基因组文库中分离出的基因的多个基因特异性引物，而不是根据已知的酶家族设计的简并引物。通过对糖苷水解酶家族9纤维素酶在元基因组DNA中的同源搜索，证明了MGSM-PCR的实用性。通过凝胶电泳法将产物显示在琼脂糖凝胶上，验证了多重聚合酶链式反应的成功。从34个土壤DNA样品中用组合多引物反应扩增出127个同源基因。多重比对显示了这些捕获序列之间广泛的序列多样性，序列同源性从26%到99.7%不等。这些结果表明，当使用多个元基因组基因特异性引物时，确实可以很容易地获得显著不同的同源基因。
A new method, termed metagenomic gene specific multi-primer PCR (MGSM-PCR), is presented that uses multiple gene specific primers derived from an isolated gene from a constructed metagenomic library rather than degenerate primers designed based on a known enzyme family. The utility of MGSM-PCR was shown by applying it to search for homologues of the glycoside hydrolase family 9 cellulase in metagenomic DNA. The success of the multiplex PCR was verified by visualizing products on an agarose gel following gel electrophoresis. A total of 127 homologous genes were amplified with combinatorial multi-primer reactions from 34 soil DNA samples. Multiple alignments revealed extensive sequence diversity among these captured sequences with sequence identity varying from 26 to 99.7%. These results indicated that significantly diverse homologous genes were indeed readily accessible when using multiple metagenomic gene specific primers.
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影响因子：
4.4
作者：
Eschenfeldt, WH;Stols, L;Donnelly, MI
通讯作者：
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影响因子：
14.9
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影响因子：
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作者：
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