Open AccessIdentification of the genus Geobacillus using genus‐specific primers, based on the 16S–23S rRNA gene internal transcribed spacer
Author(s) -
Kuisiene Nomeda,
Raugalas Juozas,
Stuknyte Milda,
Chitavichius Donaldas
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2007.00954.x
Subject(s) - internal transcribed spacer , 23s ribosomal rna , biology , 16s ribosomal rna , ribosomal rna , gene , genetics , identification (biology) , botany , rna , ribosome
The aim of this study was to develop an easy and accurate technique for the identification of the genus Geobacillus . For this purpose, Geobacillus genus‐specific primers GEOBAC (GEOBAC‐F and GEOBAC‐R) based on the 16S–23S rRNA gene internal transcribed spacer (ITS) region sequences have been designed. In total, 52 sequences from three species of the genus Geobacillus ( Geobacillus stearothermophilus , Geobacillus kaustophilus and Geobacillus lituanicus ) were examined for the design of these primers. Analysis of the sequences revealed three highly conservative regions common to these species: 5′ and 3′ end regions of 16S–23S rRNA gene ITSs and box A. Some sequences possessed two additional conservative regions – genes of tRNA Ile and tRNA Ala . These particular sequences were chosen for the construction of the primers. The designed primers targeted the gene of tRNA Ile and the 3′ end region of ITSs. This technique was validated with both the reference strains of the genus Geobacillus and the thermophilic aerobic endospore‐forming environmental isolates. Different Geobacillus species could be grouped according to the number and size of GEOBAC‐PCR products and identified on the basis of the AluI and TaqI restriction analysis of these products.