Van Camp G, Chapelle S, De Wachter R
Department of Biochemistry, University of Antwerp (UIA), Belgium.
Curr Microbiol. 1993 Sep;27(3):147-51. doi: 10.1007/BF01576012.
Published bacterial 23S ribosomal RNA sequences were aligned, and universally conserved regions flanking highly variable regions were looked for. In strategically positioned conserved regions, six oligonucleotides suitable for polymerase chain reaction (PCR) and sequencing were designed, allowing fast sequencing of four of the most variable 23S rRNA regions. Two other primers were designed for PCR amplification of nearly complete 23S rRNA genes. All these primers successfully amplified fragments of 23S rRNA genes from seven unrelated bacteria. Four primers were used to determine 938 bp of sequence for Campylobacter jejuni subsp. jejuni. These results indicate that the oligonucleotide sequences presented here are useful for PCR amplification and sequence determination of variable 23S rRNA regions for a broad variety of eubacterial species.
对已发表的细菌23S核糖体RNA序列进行比对，寻找高变区两侧的普遍保守区域。在战略定位的保守区域中，设计了六个适合聚合酶链反应（PCR）和测序的寡核苷酸，从而能够快速对四个最可变的23S rRNA区域进行测序。另外设计了两个引物用于几乎完整的23S rRNA基因的PCR扩增。所有这些引物都成功地从七种不相关细菌中扩增出了23S rRNA基因片段。使用四个引物确定了空肠弯曲菌空肠亚种938 bp的序列。这些结果表明，本文给出的寡核苷酸序列可用于多种真细菌物种可变23S rRNA区域的PCR扩增和序列测定。
