An Oligonucleotide Primer System for Amplification of the Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Genes of Bacteria of Various Taxonomic Groups |
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| Authors: | Spiridonova E M Berg I A Kolganova T V Ivanovsky R N Kuznetsov B B Tourova T P |
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| Institution: | (1) Department of Microbiology, Biological Faculty, Moscow State University, Vorob'evy gory, Moscow, 119992, Russia;(2) Russian Academy of Sciences, Bioengineering Center, pr. 60-letiya Oktyabrya 7, k. 1, Moscow, 117312, Russia;(3) Russian Academy of Sciences, Winogradsky Institute of Microbiology, pr. 60-letiya Oktyabrya 7, k. 2, Moscow, 117312, Russia |
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| Abstract: | Based on the analysis of GenBank nucleotide sequences of the cbbL and cbbM genes, coding for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC), the key enzyme of the Calvin cycle, a primer system was designed that allows fragments of these genes about 800 bp long to be PCR-amplified for various photo- and chemotrophic bacteria. The efficiency of the designed primer system in detection of RuBPC genes was demonstrated in PCR with DNA of taxonomically diverse bacteria possessing RuBPC genes with a known primary structure. Nucleotide sequences of RuBPC gene fragments of bacteria belonging to the genera Acidithiobacillus, Ectothiorhodospira, Magnetospirillum, Methylocapsa, Thioalkalispira, Rhodobacter, and Rhodospirillum were determined to be deposited with GenBank and to be translated into amino acid sequences and subjected to phylogenetic analysis. |
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| Keywords: | ribulose-1 5-bisphosphate carboxylase/oxygenase cbbL cbbM PCR oligonucleotide primers phylogenetic analysis |
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