• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Journal of Life Science
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• v.34 no.2
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• pp.86-93
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• 2024

Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.

• Korean Journal of Environmental Agriculture
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• v.40 no.4
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• pp.279-289
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• 2021

BACKGROUND: Recently, the golden apple snail, Pomacea canaliculata has been used as an environmentally-friendly weed-control agent in rice farming. Although effective for this particular style of farming, P. canaliculata can be destructive to other crops. The objective of this study was to identify overwintering as well as regional and seasonal distribution characteristics of P. canaliculata. Notably, winter is typically fatal for P. canaliculata. However, owing to increasing average global temperatures, we assessed the ability of P. canaliculata to survive through uncharacteristically warm winters. METHODS AND RESULTS: To examine the distribution and overwintering regions of P. canaliculata, We conducted a survey from April 2020 to May 2021 on environmentally-friendly rice fields, agricultural waterways, and streams in 23 cities belonging to 8 provinces. In addition, because air temperature may influence the distribution density of P. canaliculata, we analyzed the winter temperature data (http://weather.rda.go.kr). CONCLUSION(S): In 2021, overwintering of P. canaliculata (1-3 individuals/m²) was observed in the Goheung and Yeongam regions in Jeonnam. Overwintering of P. canaliculata was observed in fewer regions in 2021 than in 2020; this fact may be attributed to the lower minimum temperatures measured in 2021 (approximately 8℃ lower) than those in 2020. Our results suggest that overwintering occurs as long as overnight temperatures are ≥ -15℃, but can take place if temperatures are as low as -19℃.