Mercury bioremediation by mercury resistance transposon-mediated in situ molecular breeding.

Mercury-resistant (HgR) bacteria occur in various bacterial species from a wide variety of environmental sources. Resistance is conferred by a set of operon genes termed the mer operon. Many HgR bacteria have been isolated from diverse environments and clinical samples, and it is recognized that mer operons are often localized on transposons. Previous research reports have suggested that HgR transposons participate in the horizontal gene transfer of mer operons among bacteria. This was confirmed by a study that found that mer operons were distributed worldwide in Bacilli with dissemination of TnMERI1-like transposons. In this mini review, possible strategies for transposon-mediated in situ molecular breeding (ISMoB) of HgR bacteria in their natural habitat are discussed. In ISMoB, the target microorganisms for breeding are indigenous bacteria that are not HgR but that are dominant and robust in their respective environments. Additionally, we propose a new concept of bioremediation technology for environmental mercury pollution by applying transposon-mediated ISMoB for environmental mercury pollution control.

Development of suitable hydroponics system for phytoremediation of arsenic-contaminated water using an arsenic hyperaccumulator plant Pteris vittata.

In this study, we found that high-performance hydroponics of arsenic hyperaccumulator fern Pteris vittata is possible without any mechanical aeration system, if rhizomes of the ferns are kept over the water surface level. It was also found that very low-nutrition condition is better for root elongation of P. vittata that is an important factor of the arsenic removal from contaminated water. By the non-aeration and low-nutrition hydroponics for four months, roots of P. vittata were elongated more than 500 mm. The results of arsenate phytofiltration experiments showed that arsenic concentrations in water declined from the initial concentrations (50 µg/L, 500 µg/L, and 1000 µg/L) to lower than the detection limit (0.1 µg/L) and about 80% of arsenic removed was accumulated in the fern fronds. The improved hydroponics method for P. vittata developed in this study enables low-cost phytoremediation of arsenic-contaminated water and high-affinity removal of arsenic from water.

Mercury bioremoval by Yarrowia strains isolated from sediments of mercury-polluted estuarine water.

Difference in mercuric ion removal by resting and growing cells of two mercury-resistant yeast strains, identified as Yarrowia spp. (strains Idd1 and Idd2), were studied. Resting cells of strain Idd2 exhibited high maximum Hg(2+) removal capacity (59 mg mercury per g dry cell weight [gdw(-1)]) by adsorption than those of resting cells of strain Idd1 (32 mg gdw(-1)). The resting cells of strain Idd2 exhibited a higher Hg(2+) desorption capacity using CaCl2 (68 %) and EDTA (48 %) than strain Idd1, depicting weaker binding of Hg(2+) onto strain Idd2 unlike strain Idd1. The actively growing yeast cells showed opposite Hg removal characteristics to those of the resting cells. Strain Idd1 adsorbed less Hg(2+) from culture medium supplemented with Hg(2+) than strain Idd2. However, the growing strain Idd1 reduced and vaporized 27 % of supplemented Hg(2+) as metallic mercury (Hg(0)), while the growing strains Idd2 vaporized 15 % of the supplemented Hg(2+). These two yeast strains are potential biotechnological tools for the eventual bioremediation of polluted aquatic systems.

Evaluation of the effectiveness and salt stress of Pteris vittata in the remediation of arsenic contamination caused by tsunami sediments.

On March 11, 2011, one of the negative effects of the tsunami phenomenon that devastated the Pacific coast of the Tohoku district in Japan was the deposition of a wide range of arsenic (As) contamination to the soil. To remediate such a huge area of contamination, phytoremediation by Pteris vittata, an As-hyperaccumulator, was considered. To evaluate the efficacy of applying P. vittata to the area, the salt tolerance of P. vittata and the phytoextraction of As from soil samples were investigated. For the salt tolerance test, spore germination was considerably decreased at an NaCl level of more than 100 mM. At 200 mM, the gametophytes exhibited a morphological defect. Furthermore, the growth inhibition of P. vittata was observed with a salinity that corresponded to 66.2 mS/m of electric conductivity (EC) in the soil. A laboratory phytoremediation experiment was conducted using As-contaminated soils for 166 days. P. vittata grew and accumulated As at 264 mg/kg-DW into the shoots. Consequently, the soluble As in the soil was evidently decreased. These results showed that P. vittata was applicable to the phytoremediation of As-contaminated soil with low salinity as with the contamination caused by the 2011 tsunami.

Survival of the aerobic denitrifier Pseudomonas stutzeri strain TR2 during co-culture with activated sludge under denitrifying conditions.

The aerobic denitrifier Pseudomonas stutzeri TR2 (strain TR2) has the potential to reduce nitrous oxide emissions during the wastewater treatment process. In this application, it is important to find the best competitive survival conditions for strain TR2 in complex ecosystems. To that end, we examined co-cultures of strain TR2 with activated sludge via five passage cultures in a medium derived from treated piggery wastewater that contained a high concentration of ammonium. The results are as follows: (i) The medium supported the proliferation of strain TR2 (P. stutzeri strains) under denitrifying conditions. (ii) Nitrite was a better denitrification substrate than nitrate for TR2 survival. (iii) Strain TR2 also demonstrated strong survival even under aerobic conditions. This suggests that strain TR2 is effectively augmented to the wastewater treatment process, aiding in ammonium-nitrogen removal and reducing nitrous oxide production with a partial nitrification technique in which nitrite accumulates.

Arsenic uptake by Pteris vittata in a subarctic arsenic-contaminated agricultural field in Japan: An 8-year study.

In this study, the phytoremediation potential of tropical and subtropical arsenic (As) hyperaccumulating fern Pteris vittata in an As contaminated farmland field near an abandoned goldmine was investigated. The tested field is located in a subarctic area of northeast Japan. This study was aimed at decreasing the risk of As in the soil (water-soluble As) with nurturing the soil and respecting the plant life cycle for the sustainable phytoremediation for 8 years. The field was tilled and planted with new seedlings of the fern every spring and the grown fern was harvested every autumn. The biomass and As concentration in fronds, rhizomes and roots of the fern were analyzed separately after harvesting each year. The biomass of the fronds of P. vittata was significantly affected by the yearly change of the weather condition, but As concentration in fronds was kept at 100-150 mg/kg dry weight. The accumulated As in P. vittata was higher than that of As-hyperaccumulator fern Pteris cretica, the native fern in the field trial area. Harvested biomass of P. vittata per plant was also higher than that of P. cretica. More than 43.5 g As/154 m2 (convertible to 2.82 kg of As per hectare) was removed from the farmland field by P. vittata phytoremediation at the end of the 8-year experiment. Because of the short-term plant growth period and soil tilling process, total As in soil did not show significant depletion. However, the water-soluble As in the surface and deeper soil, which is phytoavailable and easily taken in cultivated plants, decreased to 10 µg/L (Japan Environmental Quality Standard for water-soluble As in soil) by the 8-year phytoremediation using P. vittata. These research data elucidate that the tropical and subtropical As hyperaccumulating fern, P. vittata, is applicable for As phytoremediation in the subarctic climate area.

New evidence of arsenic translocation and accumulation in Pteris vittata from real-time imaging using positron-emitting 74As tracer.

Pteris vittata is an arsenic (As) hyperaccumulator plant that accumulates a large amount of As into fronds and rhizomes (around 16,000 mg/kg in both after 16 weeks hydroponic cultivation with 30 mg/L arsenate). However, the sequence of long-distance transport of As in this hyperaccumulator plant is unclear. In this study, we used a positron-emitting tracer imaging system (PETIS) for the first time to obtain noninvasive serial images of As behavior in living plants with positron-emitting 74As-labeled tracer. We found that As kept accumulating in rhizomes as in fronds of P. vittata, whereas As was retained in roots of a non-accumulator plant Arabidopsis thaliana. Autoradiograph results of As distribution in P. vittata showed that with low As exposure, As was predominantly accumulated in young fronds and the midrib and rachis of mature fronds. Under high As exposure, As accumulation shifted from young fronds to mature fronds, especially in the margin of pinna, which resulted in necrotic symptoms, turning the marginal color to gray and then brown. Our results indicated that the function of rhizomes in P. vittata was As accumulation and the regulation of As translocation to the mature fronds to protect the young fronds under high As exposure.

Potential of aerobic denitrification by Pseudomonas stutzeri TR2 to reduce nitrous oxide emissions from wastewater treatment plants.

In contrast to most denitrifiers studied so far, Pseudomonas stutzeri TR2 produces low levels of nitrous oxide (N(2)O) even under aerobic conditions. We compared the denitrification activity of strain TR2 with those of various denitrifiers in an artificial medium that was derived from piggery wastewater. Strain TR2 exhibited strong denitrification activity and produced little N(2)O under all conditions tested. Its growth rate under denitrifying conditions was near comparable to that under aerobic conditions, showing a sharp contrast to the lower growth rates of other denitrifiers under denitrifying conditions. Strain TR2 was tolerant to toxic nitrite, even utilizing it as a good denitrification substrate. When both nitrite and N(2)O were present, strain TR2 reduced N(2)O in preference to nitrite as the denitrification substrate. This bacterial strain was readily able to adapt to denitrifying conditions by expressing the denitrification genes for cytochrome cd(1) nitrite reductase (NiR) (nirS) and nitrous oxide reductase (NoS) (nosZ). Interestingly, nosZ was constitutively expressed even under nondenitrifying, aerobic conditions, consistent with our finding that strain TR2 preferred N(2)O to nitrite. These properties of strain TR2 concerning denitrification are in sharp contrast to those of well-characterized denitrifiers. These results demonstrate that some bacterial species, such as strain TR2, have adopted a strategy for survival by preferring denitrification to oxygen respiration. The bacterium was also shown to contain the potential to reduce N(2)O emissions when applied to sewage disposal fields.

Expressing a bacterial mercuric ion binding protein in plant for phytoremediation of heavy metals.

A specific mercuric ion binding protein (MerP) originating from transposon TnMERI1 of Bacillus megaterium strain MB1 isolated from Minamata Bay displayed good adsorption capability for a variety of heavy metals. In this study, the Gram-positive MerP protein was expressed in transgenic Arabidopsis to create a model system for phytoremediation of heavy metals. Under control of an actin promoter, the transgenic Arabidpsis showed higher tolerance and accumulation capacity for mercury, cadium and lead when compared with the control plant. Results from confocal microscopy analysis also indicate that MerP was localized at the cell membrane and vesicles of plant cells. The developed transgenic plants possessing excellent metal-accumulative ability could have potential applications in decontamination of heavy metals.

Bio-oxidation of elemental mercury during growth of mercury resistant yeasts in simulated hydrosphere.

Transformation of metallic mercury (Hg°) to mercuric ion (Hg2+) in hydrosphere is the entrance of mercury cycle in water environments and leads to toxicological impact of serious global concern. Two yeast strains of Yarrowia (Idd1 and Idd2) isolated from Hg-contaminated sediments were studied for their mediating role in Hg° dissolution and oxidation. Growth of the Yarrowia cells in Hg-free liquid medium, incubated for 5 d in closed air-tight systems containing Hg°, produced extracellular polymeric substances (EPS). Approximately 230 (±5.7) ng and 120 (±6.8) ng of the dissolved Hg° were oxidized to Hg2+ by the cultures of Idd1 and Idd2, respectively, 5 day post-inoculation. Transmission electron microscopy (TEM) and X-ray energy dispersive spectrophotometry (XEDS) analysis of the EPS and cell mass revealed the presence of extracellular Hg nanoparticles, presumably HgS, as an indication of EPS-Hg complexation that is useful for Hg° dissolution and its eventual oxidation to Hg2+ by the cells. Fourier transmission infra-red (FTIR) analyses of the EPS and cell-mass during Hg-oxidation revealed that amine and carbonyl groups were used by EPS for Hg complexation. Our findings provided information about mediatory role played by Yarrowia (Idd1 and Idd2) in hydrosphere in biogeochemical cycling of Hg.

Identification of A Novel Arsenic Resistance Transposon Nested in A Mercury Resistance Transposon of Bacillus sp. MB24.

A novel TnMERI1-like transposon designated as TnMARS1 was identified from mercury resistant Bacilli isolated from Minamata Bay sediment. Two adjacent ars operon-like gene clusters, ars1 and ars2, flanked by a pair of 78-bp inverted repeat sequences, which resulted in a 13.8-kbp transposon-like fragment, were found to be sandwiched between two transposable genes of the TnMERI1-like transposon of a mercury resistant bacterium, Bacillus sp. MB24. The presence of a single transcription start site in each cluster determined by 5'-RACE suggested that both are operons. Quantitative real time RT-PCR showed that the transcription of the arsR genes contained in each operon was induced by arsenite, while arsR2 responded to arsenite more sensitively and strikingly than arsR1 did. Further, arsenic resistance complementary experiments showed that the ars2 operon conferred arsenate and arsenite resistance to an arsB-knocked out Bacillus host, while the ars1 operon only raised arsenite resistance slightly. This transposon nested in TnMARS1 was designated as TnARS1. Multi-gene cluster blast against bacteria and Bacilli whole genome sequence databases suggested that TnMARS1 is the first case of a TnMERI1-like transposon combined with an arsenic resistance transposon. The findings of this study suggested that TnMERI1-like transposons could recruit other mobile elements into its genetic structure, and subsequently cause horizontal dissemination of both mercury and arsenic resistances among Bacilli in Minamata Bay.

Comparative geochemical evaluation of toxic metals pollution and bacterial communities of industrial effluent tributary and a receiving estuary in Nigeria.

Toxic metals/metalloid contaminations of estuarine sediments due to compromised tributaries arouse significant interest in studying bacterial community that triggers natural attenuation processes. Geo-accumulation index (Igeo), contamination factor (CF), pollution load index (PLI), and Hakanson potential ecological risk index (RI) as a sum of risk factors (Er) were used to quantify toxic metal/metalloid-pollution status of Lagos Lagoon (2W) and 'Iya-Alaro' tributary (4W) sediments in comparison with pristine 'Lekki Conservation Centre' sediment (L1-B). Bacteriology of the ecosystems was based on culture-independent analyses using pyrosequencing. 2W and 4W were extremely contaminated with mercury (Igeo > 7), whereas, cadmium contamination was only observed in 4W. The two ecosystems were polluted with toxic metal based on PLI, where mercury (Er = 2900 and 1900 for 4W and 2W, respectively) posed very high ecological risks. Molecular fingerprinting revealed that Proteobacteria, Firmicutes, and Acidobacteria predominately contributed the 20 most abundant genera in the two ecosystems. The 240 and 310 species present in 2W and 4W, respectively, but absent in L1-B, thrive under the metal concentrations in the polluted hydrosphere. Whereas, the 58,000 species missing in 2W and 4W but found in L1-B would serve as indicators for systems impacted with metal eco-toxicity. Despite toxic metal pollution of the ecosystems understudied, bacterial communities play vital roles in self-recovery processes occurring in the hydrosphere.

Facilities for transcription and mobilization of an exon-less bacterial group II intron nested in transposon TnMERI1.

Transcription and mobilization facilities of a group II intron, B.me.I1, which was classified within a group IIB subclass and nested in a broad-spectrum mercury resistance transposon TnMERI1 found from the chromosome of Bacillus megaterium strain MB1 were investigated. Though B.me.I1 does not intervene in any recognizable exon gene, the splicing ability of B.me.I1 in Escherichia coli was confirmed by applying RT-PCR with RNA template transcribed by using a T7 RNA polymerase-promoter expression system. The homing activity of B.me.I1 was confirmed by using an intron-less allele fragment as a specific integration site which was cloned from Tn5084, a TnMERI1-family mercury resistance transposon. The transcription and splicing of B.me.I1 in the original host, the strain MB1, were also observed as its natural property. Primer-walking and 5' RACE analyses showed that the transcription start site and the putative promoter region of B.me.I1 were located on the antisense strand of tnpR gene of TnMERI1. Therefore, it is cleared that TnMERI1 provides a specific integration site and facilities for transcription of B.me.I1. From these results, it is considered that these genetic facilities given by the transposon TnMERI1 confer splicing and homing capability on B.me.I1, even though the group II intron is not associated with any exon gene.

Successions of bacterial community in composting cow dung wastes with or without hyperthermophilic pre-treatment.

Comparative analyses of bacterial community successions in the composting materials were done for a conventional windrow post-treatment (WPOT) process with the hyperthermophilic pre-treatment (HTPRT) and simple windrow composting (SWC; without the HTPRT). Multidimensional scaling profiles based on data of terminal restriction fragment length polymorphisms of the bacterial population in the samples of every 7 days composting material and analyses of the 16S rRNA gene-based clone library of the 7 and 21 days composting materials suggested that bacterial communities of the composting materials differed much between these two processes until the 35 days of composting, whereas that they were closely related to each other at the final composting stage (42 days of composting). Detailed phylogenetic analysis clarified that all WPOT clone libraries contained many clones of the lineages of aerobic bacteria (for example, bacilli). However, the most abundant clones retrieved from all SWC materials were affiliated with a clone cluster closely related to identified and classified members of the phylum Firmicutes that have strictly anaerobic metabolism pathways. From these results, we conclude that the HTPRT process contributed to easily establish an aerobic ecosystem from the early stage to the final stage of WPOT composting with plowing the materials only once a week.

Interactions between two MerR regulators and three operator/promoter regions in the mercury resistance module of Bacillus megaterium.

The mercury resistance module of Bacillus transposon TnMERI1 is regulated by three operator/promoter regions (O/P merB3, O/P merR1, and O/P merR2) and two regulatory proteins (MerR1 and MerR2) encoded by the module itself. To clarify the roles of MerR1 and MerR2 in the regulatory mechanism, both proteins were overexpressed and purified. MerR1 bound the regulatory regions O/P merB3 and O/P merR1, with a preference for O/P merB3 as measured on in vitro gel shift assays. However, MerR2 bound O/P merR2, as revealed by gel shift and restriction endonuclease protection assays. The transcriptional start sites of O/P merB3 and O/P merR2 were determined by rapid amplification of 5'-cDNA ends (5'-RACE) in the TnMERI1 original host, Bacillus megaterium strain MB1. Real-time reverse transcription polymerase chain reaction (RT-PCR) assays showed that O/P merB3 and O/P merR1 were induced in the presence of Hg2+ but not O/P merR2. It was concluded that MerR1 regulates O/P merB3 and O/P merR1, while MerR2 regulates O/P merR2.

Composting cattle dung wastes by using a hyperthermophilic pre-treatment process: characterization by physicochemical and molecular biological analysis.

To solve malodorous odor problems by ammonia emission in composting of cattle dung wastes, we developed an alternative composting method consisting of a hyperthrmophilic pre-treatment reactor (HTPRT) (first step) combined with a general windrow post-treatment system (WPOT) (second step). In this study, physicochemical and microbiological differences in compost materials during the HTPRT-WPOT process and a simple windrow composing process (SWC) were investigated. The HTPRT-WPOT process removed excess ammonia in the compost materials by physical ammonia stripping, and controlled the malodorous ammonia emission. The organic matter evolution index showed that the HTPRT-WPOT process also contributed to accelerate formation of humic acids in composting. Quantitative real-time PCR analyses using Bacterial-, Archaeal- and fungal-protozoan-specific primer sets showed that small subunit ribosomal RNA (SSU rRNA) gene copy numbers differed much between composting materials of these two processes. Particularly, the SSU rRNA gene copy of eukaryotic microbes (fungi-protozoa) in the HTPRT-WPOT process was much higher than in the SWC process. From these results, we conclude that the HTPRT-WPOT process has great advantages for the control of malodorous odor problems caused by ammonia emission, and for high rate of composting evaluated by the humification rate and microbial characterization of the composting materials.

Mercury removal during growth of mercury tolerant and self-aggregating Yarrowia spp.

Ecotoxicological implications of mercury (Hg) pollution of hydrosphere require effective Hg-removal strategies as antidote to the environmental problems. Mercury-tolerant yeasts, Yarrowia spp. Idd1 and Idd2 strains, were studied for intracellular accumulation and extracellular micro-precipitation of Hg during growth stage of the yeast strains. In a liquid medium containing 870 (±23.6) µg of bioavailable Hg2+, 419.0 µg Hg2+ (approx.) was taken up by the wet biomasses of the yeast strains after 48 h post-inoculation. Large portion of the adsorbed Hg was found in cell wall (approx. 49-83 %) and spheroplast (approx. 62-89 %). Negligible quantities of Hg were present in the mitochondria (0.02-0.02 %), and appreciable amount of Hg was observed in nuclei and cell debris (15.2-65.3 %) as evidence of bioaccumulation. Extracellular polymeric substances (EPS) produced by the growing Yarrowia cells was a complex of protein, carbohydrates and other substances, immobilizing 43.8 (±0.7)-58.7 (±1.0) % of initial Hg in medium as micro-precipitates, while 10.13 ± 0.4-39.2 ± 4.3 % Hg content was volatilized. Transmission electron microscopy coupled with X-ray energy dispersive spectrophotometry confirmed the cellular removal of Hg and formation of EPS-Hg complex colloids in the surrounding bulk solution as micro-precipitates in form of extracellular Hg-nanoparticles. Hg mass balance in the bio-sequestration experiment revealed excellent Hg removal (>97 %) from the medium (containing ≤16 µg ml-1 Hg2+) by the yeast strains via bioaccumulation, volatilization and micro-precipitation. The yeast strains are also effectively applicable in biological purification technology for Hg contaminated water because of their high self-aggregation activity and separatability from the aquatic environments. Graphical abstract Yarrowia species are oligotrophic marine yeasts that exhibited great potentials for mercuric ion remediation technologies, which are classified into four categories based on the process acting on the metal. These include immobilization through biosorption, compartmentation via bioaccumulation, separation from bulk solution via micro-precipitation upon EPS-Hg complex formation, and destruction that is a process to reduce the mercuric ion to metallic mercury.

Extracellular mercury sequestration by exopolymeric substances produced by Yarrowia spp.: Thermodynamics, equilibria, and kinetics studies.

Exopolymeric substances (EPS) produced by highly mercury-resistant strains of the yeast Yarrowia spp. (Idd1 and Idd2) were isolated and studied for their mercury binding potential. Excellent yield (approximately 0.3 g EPS per gram biomass) of soluble EPS in medium with 3% glucose was observed in the Yarrowia cultures 7 day post-inoculation. A gram dry weight of the EPS consists mainly of carbohydrates (0.4 g), protein (0.3-0.4 g), uronic acid (0.02 g), and nucleic acids (0.002 g). Mercury interactions with the biopolymer were measured as uptake kinetics from a simulated aquatic system and modelled with thermodynamics and calculated mass action equilibria. The EPS forms a complex with Hg2+ in water with small activation energy (≤2 kJ mol-1), achieving about 30 mg Hg2+ adsorption per gram dry weight of EPS. The adsorption models confirmed complexation of Hg2+ by the EPS via heterogeneous multilayer adsorption that obey second-order kinetics at constant rate of 4.0 and 8.1 mg g-1 min-1. The EPS used chemisorption as rate-limiting step that controls the uptake of Hg2+ from aquatic systems during micro-precipitation as bio-removal strategy. The EPS are promising biotechnological tools to design bioreactors for treatment of mercury-rich industrial wastewater.

Mercury resistance transposons in Bacilli strains from different geographical regions.

A total of 65 spore-forming mercury-resistant bacteria were isolated from natural environments worldwide in order to understand the acquisition of additional genes by and dissemination of mercury resistance transposons across related Bacilli genera by horizontal gene movement. PCR amplification using a single primer complementary to the inverted repeat sequence of TnMERI1-like transposons showed that 12 of 65 isolates had a transposon-like structure. There were four types of amplified fragments: Tn5084, Tn5085, Tn(d)MER3 (a newly identified deleted transposon-like fragment) and Tn6294 (a newly identified transposon). Tn(d)MER3 is a 3.5-kb sequence that carries a merRETPA operon with no merB or transposase genes. It is related to the mer operon of Bacillus licheniformis strain FA6-12 from Russia. DNA homology analysis shows that Tn6294 is an 8.5-kb sequence that is possibly derived from Tn(d)MER3 by integration of a TnMERI1-type transposase and resolvase genes and in addition the merR2 and merB1 genes. Bacteria harboring Tn6294 exhibited broad-spectrum mercury resistance to organomercurial compounds, although Tn6294 had only merB1 and did not have the merB2 and merB3 sequences for organomercurial lyases found in Tn5084 of B. cereus strain RC607. Strains with Tn6294 encode mercuric reductase (MerA) of less than 600 amino acids in length with a single N-terminal mercury-binding domain, whereas MerA encoded by strains MB1 and RC607 has two tandem domains. Thus, Tn(d)MER3 and Tn6294 are shorter prototypes for TnMERI1-like transposons. Identification of Tn6294 in Bacillus sp. from Taiwan and in Paenibacillus sp. from Antarctica indicates the wide horizontal dissemination of TnMERI1-like transposons across bacterial species and geographical barriers.

Participation of the recA determinant in the transposition of class II transposon mini-TnMERI1.

As an initial step to understand the mobile nature of class II mercury resistance transposon TnMERI1, the effect of the recA gene on translocation of mini-TnMERI1 was evaluated. A higher transposition frequency in the LE392 strain (2.4+/-1.2x10(-5)) than in the recA-deficient DH1 strain (1.2+/-0.8x10(-6)) indicated participation of the recA gene in mini-TnMERI1 transposition. Introduction of the recA gene into the DH1 strain complemented the transposition frequency at the same level as in LE392 and confirmed participation of the recA gene in transposition. However, treatment of cells by stress agents, including irradiation of up to 3000 Jm(-2) UV doses, did not alter the transposition frequency and suggested independence of RecA from the SOS stress response. Further analysis of transconjugants indicated participation of RecA in the resolution of the cointegrate structure of the transposon. These results suggested that RecA is a constitutive cellular factor that increases translocation of mini-TnMERI1 and may participate in dissemination of TnMERI1-like transposons.