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Genus-specific primers targeting the 16S rRNA gene for PCR detection of members of the genus Verrucosispora

文献类型： 外文期刊

作者： Xie, Qingyi 2 ; Hong, Kui 1 ; Goodfellow, Michael 3 ;

作者机构： 1.Wuhan Univ, Coll Pharm, Wuhan 430071, Peoples R China

2.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Haikou 571101, Peoples R China

3.Univ Newcastle, Sch Biol, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England

4.Hainan Univ, Coll Agron, Haikou 570228, Peoples R China

关键词： Verrucosispora;Genus-specific primers;Marine samples;Environmental clones

期刊名称：ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY （ 影响因子：2.271； 五年影响因子：2.707 ）

ISSN： 0003-6072

年卷期： 2011 年 100 卷 1 期

页码：

收录情况： SCI

摘要： Little is known about the genus Verrucosispora though it does contain organisms which produce novel antibiotics. A set of genus-specific oligonucleotide primers was generated to gain an insight into the presence, distribution and taxonomic diversity of members of this genus in diverse samples taken from marine habitats. In silico and pure culture studies showed that the primers matched perfectly with target sequences of the 16S rRNA genes of representatives of the genus Verrucosispora. The primers, designated S-G-Verr-0195-a-S-20 and S-G-Verr-1152-a-A-18, amplified an a parts per thousand 960 bp stretch of the 16S rRNA genes of Verrucosispora strains but not those of representatives of other genera classified in the family Micromonosporaceae. Genus-specific amplicons were detected from 17 out of 20 community DNA samples prepared from diverse marine sediments and coastal soils. Phylogenetic analysis of over 40% of clones derived from five of the samples indicated they belonged to novel Verrucosispora species. The primers were also used to confirm the identity of Verrucosispora-like strains isolated from two of the environmental samples. The primers can be used to facilitate the isolation of novel Verrucosispora strains by allowing prescreening of environmental samples and the subsequent identification of verrucosisporae on selective isolation plates. For this purpose, a novel medium facilitating the recovery of Verrucosispora strains was formulated and used to recover novel isolates validated using the novel PCR primers. This medium may be useful as the basis for development of a selective medium.

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