caa a22 4500 397552416 CHVBK 20180308164753.0 cr unu---uuuuu 161202e199607 xx s 000 0 eng 10.1111/j.1574-6941.1996.tb00315.x doi (NATIONALLICENCE)oxford-10.1111/j.1574-6941.1996.tb00315.x The metabolic pathway of 2,4-dichlorophenoxyacetic acid degradation involves different families of tfdA and tfdB genes according to PCR-RFLP analysis [Elektronische Daten] [Tatiana Vallaeys, Roberta R. Fulthorpe, Alice M. Wright, Guy Soulas] Twenty-five 2,4-dichlorophenoxyacetic acid (2,4-D) degrading bacteria from geographically diverse locations and presenting various degrees of similarity or no similarity to the tfdA and tfdB genes from Alcaligenes eutrophus JMP134 were analysed by PCR-RFLP (restriction length fragment polymorphism). Primers for the 2,4-D etherase gene were derived by sequence alignment of the tfdA genes from A. eutrophus JMP134 and Burkholderia sp. RASC. Primers for the 2,4-dichlorophenolhydroxylase gene were based on the tfdB gene sequence from A. eutrophus JMP134 by taking codon degeneration and variations in amino acid residue sequences into consideration. PCR amplification using the tfdA primer set produced fragments of 0.3 kb from 17 strains which showed varying degrees of similarity to the tfdA gene probe from A. eutrophus JMP134. Significant variations in the gene sequences were confirmed by PCR-RFLP analysis. DNA amplification using the tfdB primer set produced a 1.1 kb fragment from 19 strains. Amongst them, two did not show any similarity to the tfdB gene probe. The size and restriction pattern of the products obtained from A. eutrophus JMP134 were in accordance with the expected size calculated from the A. eutrophus tfdA and tfdB gene sequence and their theoretical PCR-RFLP patterns. Some strains which did not amplify using the tfdA primer set did however amplify with the tfdB primer set. These results suggest the independent evolution of these two genes in the construction of the 2,4-D metabolic pathway. Our tfdA and tfdB primer sets could be used for the detection of similar sequences in bacteria and soils. Moreover, PCR-RFLP patterns could also be used to select subsets of strains for sequencing to study the phylogeny of the tfdA and tfdB genes. © 1996 Federation of European Microbiological Societies. Published by Elsevier Science B.V. Alcaligenes eutrophus JMP134 nationallicence Burkholderia sp. RASC nationallicence PCR-primer nationallicence tfdA gene nationallicence tfdB gene nationallicence 2,4-Dichlorophenolhydroxylase nationallicence Alpha-ketoglutarate dependent dioxygenase nationallicence Vallaeys Tatiana Centre de Microbiologie du Sol et de l'Environnement, Laboratoire de Microbiologie des Sols, B.V. 1540, 21034 Dijon, France aut Fulthorpe Roberta R. Center for Microbial Ecology, Michigan State University, East-Lansing MI, USA aut Wright Alice M. Center for Microbial Ecology, Michigan State University, East-Lansing MI, USA aut Soulas Guy Centre de Microbiologie du Sol et de l'Environnement, Laboratoire de Microbiologie des Sols, B.V. 1540, 21034 Dijon, France aut FEMS Microbiology Ecology Blackwell Publishing Ltd 20/3(1996-07), 163-172 20:3<163 1996 20 femsec https://doi.org/10.1111/j.1574-6941.1996.tb00315.x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1111/j.1574-6941.1996.tb00315.x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vallaeys Tatiana Centre de Microbiologie du Sol et de l'Environnement, Laboratoire de Microbiologie des Sols, B.V. 1540, 21034 Dijon, France aut NATIONALLICENCE 700 1- Fulthorpe Roberta R. Center for Microbial Ecology, Michigan State University, East-Lansing MI, USA aut NATIONALLICENCE 700 1- Wright Alice M. Center for Microbial Ecology, Michigan State University, East-Lansing MI, USA aut NATIONALLICENCE 700 1- Soulas Guy Centre de Microbiologie du Sol et de l'Environnement, Laboratoire de Microbiologie des Sols, B.V. 1540, 21034 Dijon, France aut NATIONALLICENCE 773 0- FEMS Microbiology Ecology Blackwell Publishing Ltd 20/3(1996-07), 163-172 20:3<163 1996 20 femsec Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford