National Bureau of Agriculturally Important Microorganisms, Kusmaur, Mau Nath Bhanjan, UP, India.
Antonie Van Leeuwenhoek. 2011 Feb;99(2):283-96. doi: 10.1007/s10482-010-9487-4. Epub 2010 Aug 10.
Restriction fragment length analysis of PCR amplified 16S rDNA with AluI revealed the presence of a 265 bp fragment in all species of Bacillus with the exception of B. cereus and B. thuringiensis, which contains two restriction sites within this fragment which results in three smaller fragments totalling to 265 bp. Some distant species of Bacillus with no evidence of this fragment could be delineated into other genera based on phenotypic and genotypic parameters. BLAST search for homologous sequences of individual species revealed that it is a highly conserved region. Multiple alignment of the fragment suggests that a region between 160 and 265 bp of the 265 bp fragment was a hypervariable region and were highly species-specific. A set of primers was designed for amplification of this hypervariable region. Partial sequencing of the hypervariable region within the 265 bp fragment seems an index for identification of Bacillus species.
PCR 扩增 16S rDNA 经 AluI 限制片段长度分析显示，除了含有该片段内两个限制位点导致产生三个总计 265bp 小片段的蜡样芽孢杆菌和苏云金芽孢杆菌外，所有芽孢杆菌属的种均存在 265bp 片段。一些没有该片段证据的芽孢杆菌远缘种可根据表型和基因型参数划分为其他属。对单个种同源序列的 BLAST 搜索表明，这是一个高度保守的区域。该片段的多重比对表明，265bp 片段的 160bp 至 265bp 之间的一个区域是一个高变区，具有高度的种特异性。设计了一组引物用于扩增这个高变区。265bp 片段内高变区的部分测序似乎是鉴定芽孢杆菌种的一个指标。
