five to 99. 3% sequence identity with the PhnAc sequences iden tified to date from pure cultures. Phylogenetic analysis of PhnAc like sequences retrieved from sediment libraries and the ones from pure cultures revealed two clades, one particular containing phenanthrene dioxygenase sequences from your isolates Burkholderia sp. strains Cs1 four, Ch1 1, Ch3 five and Eh1 one and a second one with PhnAc from A. faecalis AFK2, Representatives from the two clades had been observed while in the Ac OR04 library. However, all Ac SC04 and Ac MS05 PhnAc like sequences clustered inside of the Burkholderia clade, though all Ac GR06 PhnAc like gene fragments clustered inside the A. faecalis AFK2 clade, NahAc style sequences formed two distinct, highly sup ported groups, 1 of them integrated sequences from Pseudomonas stutzeri strain AN10, Pseudomonas aeruginosa PAK1 and Pseudomonas balearica SP1402 as well as other clade included sequences belonging to P.
this article putida strains NCIB 9816 four, G7, OUS82 and also other related Pseudomonas isolates, Each groups incorporate sequences from marine isolates and have been described previously as AN10 and C18 groups respectively, Only sequences through the C18 cluster were detected in coastal sediments. This isn’t sudden since the AN10 group was most likely not targeted by the primers used within this function, Development and analysis of phnA1 like ARHD gene libraries A primer set was intended to target phnA1 like ARHD sequences from bacteria belonging for the genus Cycloclas ticus. This primer set effectively amplified a 500 bp frag ment from Cycloclasticus pugetii PS one, which was confirmed being a phnA1 gene fragment by sequencing.
Amplifications with this primer set also resulted in merchandise in the anticipated size in samples MP04, GR06, selleck inhibitor AR06, SC04, OR04 and OR05, Samples PF05, MS05, CR05, OR06, EM06 and OL06 weren’t examined with this particular primer set. The amplification products of samples SC04 and OR05 had been cloned and analyzed by RFLP examination together with the RsaI restriction endonuclease. As every one of the analyzed clones showed restriction patterns identi cal towards the 1 made from the amplification solution of C.