• Title/Summary/Keyword: conserved domain database

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Galectin-1 from redlip mullet Liza haematocheilia: identification, immune responses, and functional characterization as pattern recognition receptors (PRRs) in host immune defense system

  • Chaehyeon Lim;Hyukjae Kwon;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.25 no.11
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    • pp.559-571
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    • 2022
  • Galectins, a family of ß-galactoside-binding lectins, have emerged as soluble mediators in infected cells and pattern recognition receptors (PRRs) responsible for evoking and regulating innate immunity. The present study aimed to evaluate the role of galectin-1 in the host immune response of redlip mullet (Liza haematocheilia). We established a cDNA database for redlip mullet, and the cDNA sequence of galectin-1 (LhGal-1) was characterized. In silico analysis was performed, and the spatial and temporal expression patterns in gills and blood in response to lipopolysaccharide polyinosinic:polycytidylic acid, and Lactococcus garvieae were estimated via quantitative real-time PCR. Functional assays were conducted using recombinant protein to investigate carbohydrate binding, bacterial binding, and bacterial agglutination activity. LhGal-1 was composed of 135 amino acids. Conserved motifs (H-NPR, -N- and -W-E-R) within the carbohydrate recognition domain were found in LhGal-1. The tissue distribution revealed that the healthy stomach expressed high levels of LhGal-1. The temporal monitoring of LhGal-1 mRNA expression in the gill and blood showed its significant upregulation in response to immune challenges with different stimulants. rLhGal-1 exhibited binding activity in response to carbohydrates and bacteria. Moreover, the agglutination of rLhGal-1 against Escherichia coli was observed. Collectively, our findings suggest that LhGal-1 may function as a PRR in redlip mullet. Furthermore, LhGal-1 can be considered a significant gene to play a protective role in redlip mullet immune system.

Bacterial Diversity in the Mud Flat of Sunchon Bay, Chunnam Provice, by 16S rRNA Gene Analysis (16S rRNA 유전자 분석에 의한 전남 순천만 갯벌의 세균 다양성)

  • 이명숙;홍순규;이동훈;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.137-144
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    • 2001
  • In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.

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Isolation, characterization and expression of transcription factor ScDREB2 from wild, commercial and interspecific hybrid sugarcane in salinity condition

  • Chanprame, Sontichai;Promkhlibnil, Tanawan;Suwanno, Sakulrat;Laksana, Chanakan
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.97-105
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    • 2019
  • Dehydration Responsive Element Binding (DREB) gene is one of the essential transcription factors plants use for responding to stress conditions including salinity, drought, and cold stress. The purpose of this study was to isolate the full length and characterize the DREB gene from three different genotypes of sugarcane, wild, commercial cultivar, and interspecific hybrid sugarcane. The length of the gene, designated ScDREB was 789 bp, and coding for a putative polypeptide of 262 amino acid residues. Sequences of the gene were submitted to the GenBank database with accession numbers of KX280722.1, KX280721.1, and KX280719.1 for wild sugarcane, commercial cultivar (KPS94-13), and interspecific hybrid (Biotec2), respectively. In silico characterization indicated that the deduced polypeptide contains a putative nuclear localization signal (NLS) sequence, and a conserved AP2/ERF domain of the DREB family, at 82-140 amino residues. Based on multiple sequence alignment, sequences of the gene from the three sugarcane genotypes were classified in the DREB2 group. Gene expression analysis indicated, that ScDREB2 expression pattern in tested sugarcane was up-regulated by salt stress. When the plants were under 100 mM NaCl stress, relative expressions of the gene in leaves was higher than those in roots. In contrast, under 200 mM NaCl stress, relative expressions of the gene in roots was higher than those in leaves. This is the first report on cloning the full length and characterization, of ScDREB2 gene of sugarcane. Results indicate that ScDREB2 is highly responsive to salt stress.

Transcription Factor for Gene Function Analysis in Maize (옥수수 유전자 기능 분석을 위한 전사인자의 이해)

  • Moon, Jun-Cheol;Kim, Jae Yoon;Baek, Seong-Bum;Kwon, Young-Up;Song, Kitae;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.263-281
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    • 2014
  • Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3/VP1, AP2/EREBP, ARF, ARID, AS2, AUX/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2-YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.

Molecular genetic analysis of phytochelatin synthase genes in Arabidopsis

  • Ha, Suk-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 2002.04a
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    • pp.62-72
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    • 2002
  • This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined

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