PCR-mediated detection of the chemolithotrophic bacterium Thiobacillus cuprinus using 23S rDNA- and 16S/23S intergenic spacer region-targeted oligonucleotide primers.
Bioleaching is carried out by chemolithotrophic microorganisms, most of them belonging to the genera Thiobacillus and Leptospirillum. The role of the mixotrophic species T. cuprinus in this process is controversial, since its ecological study applying classical detection techniques to natural or industrial environments is very difficult. For this reason, we have developed an alternative method based on PCR-mediated detection using specific oligonucleotide primers that target variable regions of the 23S rRNA coding gene and of the 16S/23S intergenic spacer region. Specificity and sensitivity of PCR amplifications performed with both kinds of primers were studied.
Moreira
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Centro de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain, dmoreira@mvax.cbm.uam.es
Amils
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D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D013855:Thiobacillus / Q000235:genetics* / Q000302:isolation & purification*
D017931:DNA Primers; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S