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The Heavy-Metal Resistance Determinant of Newly Isolated Bacterium from a Nickel-Contaminated Soil in Southwest Slovakia

机译:西南斯洛伐克镍污染土壤中新分离细菌的重金属抗性决定因子

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摘要

A bacterial isolate MR-CH-I2 [KC809939] isolated from soil contaminated mainly by high nickel concentrations in southwest Slovakia was previously found carrying nccA-like heavy-metal resistance determinant, marked as MR-CH-I2-HMR [KF218096]. According to phylogenetic analysis of short (696 bp) 16S rDNA (16S rRNA) sequences this bacterium was tentatively assigned to Uncultured beta proteobacterium clone GC0AA7ZA05PP1 [JQ913301]. nccA-like gene product was on the same base of its partial (581 bp) sequences tentatively assigned to CzcA family heavy metal efflux pump [YP_001899332] from Ralstonia picketii 12J with 99% similarity. In this study the bacterium MR-CH-I2 and its heavy-metal resistance determinant were more precisely identified. This bacterial isolate was on the base of phylogenetic analysis of almost the whole (1,500 bp) 16S rDNA (16S rRNA) sequence, MR-CH-I2 [MF102046], and sequence for gyrB gene and its product respectively, MR-CH-I2-gyrB [MF134666], assigned to R. picketii 12J [CP001068] with 99 and 100% similarities, respectively. In addition, the whole nccA-like heavy-metal resistance gene sequence (3,192 bp), marked as MR-CH-I2-nccA [KR476581], was obtained and on the base of phylogenetic analysis its assignment was confirmed to MULTISPECIES: cation efflux system protein CzcA [WP_004635342] from Burkholderiaceae with 98% similarity. Furthermore, although the bacterium carried one high molecular plasmid of about 50 kb in size, nccA-like gene was not located on this plasmid. Finally, the results from RT-PCR analysis showed that MR-CH-I2-nccA gene was significantly induced only by the addition of nickel.
机译:从西南斯洛伐克西南斯洛伐克的高镍浓度污染的细菌分离为MR-CH-I2 [KC809939]污染的土壤中污染的污染物,载有NCCA样重金属阻力决定簇,标记为MR-CH-I2-HMR [KF218096]。根据短(696bp)16s rdna(16s rRNA)序列的系统发育分析,该细菌暂时分配给未培养的β蛋白克隆GcO 87za05pp1 [JQ913301]。 NCCA样基因产物在其部分(581bp)序列的相同碱基上,暂定于Czca Family Mevery Meflux泵[YP_001899332]来自Ralstonia Picketii 12j,其相似度为99%。在该研究中,更精确地鉴定了细菌MR-CH-I2及其重金属抗性决定簇。这种细菌分离物在几乎全部(1,500bp)16s rdNA(16s rRNA)序列,MR-CH-I2 [MF102046]和序列的序列中,SMR-CH-I2的序列分别是血管发育分析,MR-CH-I2及其产物的序列-GyRB [MF134666]分配给R. PICKETII 12J [CP001068],分别具有99和100%相似之处。此外,获得了标记为MR-CH-I2-NCCA [KR476581]的全NCCA的重金属抗性基因序列(3,192bp),并在系统发育分析的基础上证实了其作用:阳离子流动来自Burkholderiaceae的系统蛋白CZCA [WP_004635342,其相似度为98%。此外,虽然细菌载为大小约50kb的高分子质粒,但NCCA样基因未位于该质粒上。最后,RT-PCR分析结果表明,仅通过加入镍显着诱导MR-CH-I2-NCCA基因。

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