• 한국동물학회:학술대회논문집
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• 한국동물학회 1997년도 한국생물과학협회 학술발표대회
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• pp.286.2-286
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• 1997

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• Animal cells and systems
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• 제9권4호
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• pp.205-209
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• 2005

A rapid, simple and efficient method to extract RNA from the adult polyps of a soft coral, marine cnidarian, Scleronephthya gracillimum $(K\ddot{u}kenthal)$; was developed in this study. The highest yield and purity of RNA was obtained with the lysis solution containing 35 mM EDTA, 0.7 M LiCl, 7.0% SDS, and 200 mM Tris-Cl (pH 9.0). Approximately $40{\mu}g$ of total RNA was extracted from 200 mg of liquid nitrogen-pulverized polyp tissue. The ratio of absorbance at 260 nm and 280 nm ranged from 1.8 to 2.0. The results of the reverse transcription polymerase chain reaction (RTPCR) with ${\beta}-actin$ gene specific primers and Northern blot analysis using the same gene probe revealed that the RNA extracted by our method had high quality, and was sufficient for subsequent molecular biological analyses. This method was effective for RNA extraction from other soft coral species which belong to the genus Dendronephthya.

• 한국환경성돌연변이발암원학회지
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• 제19권1호
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• pp.28-33
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• 1999

The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

• 한국양식학회지
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• 제16권2호
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• pp.124-127
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• 2003

Our previous studies of both microarray analysis in channel catfish muscle gene expression of 2 different ages and channel catfish muscle expressed sequence tag profiles demonstrated parvalbumin beta is one of the highly expressed muscle transcriptome. We have cloned and sequenced complementary DNA encoding the channel catfish parvalbumin which encode 109 amino acids. The deduced amino acid sequences of the catfish parvalbumin are highly conserved with those cloned from other teleosts. The availability of the catfish parvalbumin provides the opportunity of studying fish epitopes.

• Journal of Ecology and Environment
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• 제38권4호
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• pp.397-403
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• 2015

Rice (Oryza sativa L.) is an important feeding crop in Asia, and utilization of genetically modified (GM) rice is highly demanding. For co-existence of GM rice and non-GM rice, the proper confinement measures should be provided. Thus, we surveyed gene flow from herbicide resistant GM rice to the conventional rice cultivars in the field tests. Gene flow frequency decreased with increasing distance between the pollen donor and recipients and did not exceed more than 1% even at the nearest distance. In single recipient model plot, a maximum gene flow frequency was observed at the shortest distance and hybrid was detected up to 12 m from the pollen donor. The direction of gene was coincided with the dominant wind direction. Gene flow assessment to multiple recipient plots was conducted under the high raining season by chance, and abrupt decline of gene flow frequency and maximum distance were resulted. According to the survey results, current regulation for isolation distance is reasonable for environmental safety or for general crop production. However, we suggest an alternative measure for GM rice cultivation that should be supplemented to overcome the out of estimation and in the environment asking higher security levels.

• 한국환경성돌연변이발암원학회지
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• 제16권2호
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• pp.77-82
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• 1996

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

• Food Science and Biotechnology
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• 제16권2호
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• pp.294-298
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• 2007

The actinidin (EC 3.4.22. 14) found in kiwifruit is a cysteine protease. In order to obtain the actinidin gene from the Chinese wild kiwifruit, primers were designed on the basis of the actinidin gene of Actinidia deliciosa, the New Zealand kiwifruit. The 1.2 kb DNA fragment was acquired from the total RNAs of Chinese wild kiwifruit via reverse transcription polymerase chain reaction (RT-PCR), and its DNA sequence was analyzed. Its sequence was determined to share 98.4% homology with the actinidin gene of A. deliciosa. In order to verity the actinidin gene isolated from the Chinese wild kiwifruit in Escherichia coli, the mature gene was amplified via PCR and expressed in E. coli under the control of the T7lac promoter. The actinidin was expressed in E. coli as inclusion bodies, which were solubilized with urea and refolded. The protease activity of the refolded protein was approximately twice as high as that of E. coli BL2l (DE3).

• 한국균학회지
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• 제26권3호통권86호
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• pp.314-320
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• 1998

mRNA의 differential display방법에 의해 여름느타리 자실체의 상처 또는 자외선 처리시 발현되는 약 0.4kb의 cDNA 단편을 분리하였다. 이 cDNA 단편의 염기서열 분석결과 세포분열 촉진에 관여하는 cdc2-related protein kinase gene과 상당부분 유사성을 보였으며 RT-PCR 방법을 이용한 분리 유전자의 발현 실험을 통해 이 유전자가 정상 생장 환경에서는 갓, 대, 균사 등 모든 부위에서 기본적인 발현상태를 유지하고 있으며 기계적 상처 또는 자외선 처리에 의해 그 발현이 증폭됨을 확인하였다. 이러한 결과를 통해 향후 분리된 유전자의 연구를 통한 버섯 병 방어 관련 신호 전달 체계 분석에 대한 가능성을 검토해 보았다.

• 한국미생물·생명공학회지
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• 제32권3호
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• pp.282-285
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• 2004

The polyene antibiotics including nystatin, pimaricin, amphotericin and candicidin are a family of most promising antifungal polyketide compounds, typically produced by rare actinomycetes species. The biosynthetic gene clusters for these polyenes have been previously investigated, revealing the presence of highly homologous biosynthetic genes among polyene-producers such as polyketide synthase (PKS) and cytochrome P450 hydroxylase (CYP) genes. Based on amino acid sequence alignment among actinomycetes CYP genes, the highly-conserved regions specific for only polyene CYP genes were identified and chosen for degenerate PCR primers, followed by the PCR-screening with various actinomycetes genomic DNAs. Among tested several polyene non-producing actinomycetes strains, Pseudonorcardia autotrophica strain was selected based on the presence of PCR product with polyene-specific CYP gene primers, and then confirmed to contain a cryptic novel polyene hydroxylase gene in the chromosome. These results suggest that the polyene-specific hydroxylase gene PCR should be an efficient way of screening and isolating potentially-valuable cryptic polyene antibiotic biosynthetic genes from various microorganisms including rare actinomycetes.

• 한국초지조사료학회지
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• 제21권3호
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• pp.145-150
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• 2001

This study was conducted to obtain the transformed birdsfoot trefoil (Lotus corniculatus L.) plants with BcHSP17.6 gene using Agrobacterium turnefaciens LBA4404 and we confirmed transformed gene from the regenerated birdsfoot trefoil plants. The expression vector, pBKH4 vector, harboring BcHSP17.6 gene was used for production of transgenic birdsfoot trefoil plants. The callus of birdsfoot trefoil was cocultivated with Agrobacteriurn turnefaciens and transformed calli were selected on kanamycin-containing SH-kc medium to regenerate into plants. The transformed birdsfoot trefoil plants were produced 4 momths after cultivation on BOi2Y medium. The transgenic birdsfoot trefoil plants were analyzed by isolation of genomic DNA and genomic Southern hybridization using a -32P labelled BcHSPl7.6 fragments. (Key words : Birdsfoot trefoil, Transgenic plant. BcHSP17.6 gene, Callus induction, Plant regeneration)