• 원예과학기술지
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• 제31권3호
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• pp.299-307
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• 2013

A radish (Raphanus sativus L.) polygalacturonase-inhibiting protein (PGIP) gene was cloned and compared to the PGIP gene (BrPGIP2) from Chinese cabbage (Brassica rapa ssp. pekinensis) in order to gain more information on controlling a disease and improving produce quality. To clone the radish PGIP gene, primers were designed based on conserved sequences of two PGIP genes (BnPGIP1 and BnPGIP2) from rape (B. napus L. ssp. oleifera), Chinese cabbage and Arabidopsis thaliana. PCR cloning was performed with cDNA from the stigma of radish 'Daejinyeoreum' as a template to confirm DNA fragments which were about 600 base pair in size. Sequence analysis revealed 84.1% homology with BrPGIP2 and 70.1% with BnPGIP1. DNA walking was conducted to confirm the open reading frame of 972 bp, and the gene was named RsPGIP1. RsPGIP1 consisting with 323 amino acids (aa) has a high leucine content (54/323) and contains 10 leucine-rich repeat domains, as do most BrPGIPs of Chinese cabbage. The gene expression of RsPGIP1 was induced by abiotic stresses and methyl jasmonate. It showed enrichment in the stigma and the primary root than a leaf. Cloning RsPGIP1 will aid to further apply practices on postharvest quality maintenance and disease control of the root.

• 한국미생물·생명공학회지
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• 제26권2호
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• pp.110-116
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• 1998

몇 가지 그람양성 세균에서 secY 유전자가 함유된 operon의 구성이 rplO(L15)-secY-adk라는 결과를 토대로 L15와 adk 유전자일부를 primer로 제작하여 Streptomyces lividans TK24에서 secY 유전자가 함유된 1.8kb 단편을 PCR로 증폭하여 얻은 후 secY 유전자를 cloning하였다. 전 단편을 sequencing하여 추론한 아미노산으로 상동성을 조사해 본 결과 Escherichia coli, Bacillus subtilis, Micrococcus luteus, Bacillus licheniformis, Staphylococcus carnosus, Brevibacterium flavum, Streptomyces scabies의 SecY와 각각 46%, 43%, 57%, 44%, 42%, 56%, 90%의 유사성을 보이는 것으로 나타났으며 SecY의 소수성 profile 또한 서로 유사하고 10개의 membrane spanning segment를 동일하게 가지는 것으로 나타났다. 유전자 구조도 다른 그람양성균에서와 같이 L15-SecY-Adk순 이였으며 secY 유전자의 종결코돈과 adk 유전자의 개시코돈이 한 염기를 공유하는 상태의 translational coupling 구조를 하고 있었다. 이와 같이 단백질분비에 관여하는 유전자와 ribosome 구성요소, 그리고 ATP 합성에 관여하는 요소는 세포성장에 상호 연관 작용이 있는 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.277-284
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• 1993

세계적으로 중요한 열대풍토병 병원체의 하나인 열대리슈마니아의 핵형에 영향을 줄 것으로 기대되는 인자와 그 효과를 관찰하고자 하였다. 토기의 혈액을 포함한 N.N.N. 배지에서 유지하고 있는 열대리슈마니아(Leishmania major)의 promastigote를 열쇽. 약제 첨가 자외선 조사 및 감마선을 이용한 방사선 조사를 여러 가지 방법으로 시행하고 주기변동전기영동(pulsed field gradient gel electrophoresis)을 이용하여 핵형의 변동 여부를 관찰하였다 그 결과 여러 방식에 의한 열쇽과 약제 처리 및 자외선 조사에 따른 핵형의 변화가 없었다 그러나 방사선 조사군에서는 50 Gy 이상 조사한 군에서 1 mesa base pair(Hb) 크기에 있는 염색체부터 소실되기 시작하여 방사선 조사량이 증가함에 따라서 250-500 Kb의 작은 염색체도 파괴되어 500 Gy 이상 군에서는 뚜렷한 염색체 분획이 없이 젤 하단 200 Kb 크기 아래 부분에 몰려 있었다. 이러한 소견은 방사선에 의하여 염색체가 불규칙하게 파손된 것을 의미한다고 하겠다. 방사선을 300 Gy까지 조사한 충체는 계대 배양이 가능하였고. 이들은 원래의 핵형을 유지하였다. 방사선조사 후에 배양된 충체는 염색체가 파괴되지 않았거나 부분적인 손상 후에 DNA 재결합에 의해 원상회복된 것으로 판단된다. 열대리슈마니아의 핵형은 일시적인 자극에 의하여 쉽게 변형되지 않는 안정된 것임을 확인하였다.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.651-656
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• 2013

Human schistosomiasis caused by Schistosoma japonicum and Schistosoma mekongi is a chronic and debilitating helminthic disease still prevalent in several countries of Asia. Due to morphological similarities of cercariae and eggs of these 2 species, microscopic differentiation is difficult. High resolution melting (HRM) real-time PCR is developed as an alternative tool for the detection and differentiation of these 2 species. A primer pair was designed for targeting the 18S ribosomal RNA gene to generate PCR products of 156 base pairs for both species. The melting points of S. japonicum and S. mekongi PCR products were $84.5{\pm}0.07^{\circ}C$ and $85.7{\pm}0.07^{\circ}C$, respectively. The method permits amplification from a single cercaria or an egg. The HRM real-time PCR is a rapid and simple tool for differentiation of S. japonicum and S. mekongi in the intermediate and final hosts.

• 대한예방의학회:학술대회논문집
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• 대한예방의학회 1999년도 제51차 추계 학술대회 연제집
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• pp.1-15
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• 1999

In a case-control study to evaluate the factors involved in the development of hepatocellular carcinoma, polymorphisms of the p53 gene were compared in 68 cases mostly infected with hepatitis C virus (HCV) and 68 controls matched for sex and age: DNA from peripheral blood leukocytes was analyzed by the polymerase chain reaction-single strand conformation polymorphism method and direct sequencing. Polymorphisms analyzed were those in exon 4 (CCC vs. CGC, Pro vs. Arg at codon 72, Al allele vs. A2 allele), intron 2 (C vs. G at nucleotide 38, Al vs. A2), intron 3 (C vs. A at nucleotide 65, Al vs. A2; absence and presence of 16 base pair repeat at nucleotides 24 to 39, Al vs. A2), intron 6 (A vs. G at nucleotide 62, Al vs. A2) and intron 7 (C and T vs. T and G at nucleotides 72 and 92, Al vs. A2). A significantly higher frequency of the allele for CCC (Pro, Al) at codon 72 of exon 4 was found in cases (39%) than in controls (26%) (p<0.05). Highly significant linkage of the polymorphisms in exon 4, intron 2, intron 3 and intron 7, and between the intron 3-16 bp duplication and polymorphism in intron 6 also was found. Matched Fair analysis showed significantly higher frequencies of certain haplotypes (1-1-1-1-2-2 or 1-1-2-1-2-1 for exon 4, intron 2, intron 3, the intron 3-16 bp duplication, intron 6 and intron 7) in cases than in controls (p=0.014, OR=2.27, 95% CI= 1.08-5.12). No preference of specific p53 polymorphisms for specific HCV genotype was detected. These findings suggest that in hepatocarcinogenesis mainly due to HCV infection, genetic factors may be involved and that genetic markers can serve as predictors of a high-risk group for hepatocarcinogenesis.

• 한국약용작물학회지
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• 제26권1호
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• pp.26-31
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• 2018

Background: Angelica gigas, A. sinensis, and A. acutiloba are commercially important in the herbal medicine market, and among them, A. gigas has the highest economic value and price. However, their similar morphological traits are often used for fraud. Despite their importance in herbal medicine, recognition of the differences between Angelica species is currently inadequate. Methods and Results: A multiplex polymerase chain reaction (PCR) method was developed for direct detection and identification of A. gigas, A. sinensis, and A. acutiloba. The gene for the distinction of species was targeted at ITS in the nucleus and trnC-petN gene in chloroplasts. The optimized multiplex PCR in the present study utilized each Angelica species-specific primer pairs. Each primer pair yielded products of 229 base pairs (bp) for A. gigas, 53 bp for A. sinensis, 170 bp for A. acutiloba. Additionally non-specific PCR products were not detected in similar species by species-specific primers. Conclusions: In the present study, a multiplex-PCR assay, successfully assessed the authenticity of Angelica species (A. gigas, A. sinensis, and A. acutiloba). and whole genome amplification (WGA) was performed after DNA extraction to identify, the species in the product. The detection method of raw materials developed in the present study could be applied to herbal medicine and health functional food management.

• Journal of Microbiology
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• 제42권3호
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• pp.188-193
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• 2004

Xanthobacter flavus strain UE15 was isolated in wastewater obtained from the Ulsan industrial complex, Korea. This strain functions as a 1,2-dichloroethane (1,2-DCA) degrader, via a mechanism of hydrolytic dechlorination, under aerobic conditions. The UE15 strain was also capable of dechlorinating other chloroaliphatics such as 2-chloroacetic acid and 2-chloropropionic acid. The dhlA gene encoding 1,2-DCA dechlorinase was cloned from the genomic DNA of the UE15 strain, and its nucleotide sequence was determined to consist of 933 base pairs. The deduced amino acid sequence of the DhlA dechlorinase exhibited 100％ homology with the corresponding enzyme from X. autotrophicus GJ10, but only 27 to 29％ homology with the corresponding enzymes from Rhodococcus rhodochrous, Pseudomonas pavonaceae, and Mycobacterium sp. strain GP1, which all dechlorinate haloalkane compounds. The UE15 strain has an ORF1 (1,356 bp) downstream from the dhlA gene. The OFR1 shows 99％ amino acid sequence homology with the transposase reported from X. autotrophicus GJ10. The transposase gene was not found in the vicinity of the dhlA in the GJ10 strain, but rather beside the dhlB gene coding for haloacid dechlorinase. The dhlA and dhlB genes were confirmed to be located at separate chromosomal loci in the Xanthobacter flavus UE15 strain as well as in X. autotrophicus GJ10. The dhlA and transposase the UE15 strain were found to be parenthesized by a pair of insertion sequences, 181247, which were also found on both sides of the transposase gene in the GJ10 strain. This unique structure of the dhlA gene organization in X. flavus strain UE15 suggested that the dechlorinase gene, dhlA, is transferred with the help of the transposase gene.

• 생명과학회지
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• 제21권5호
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• pp.626-630
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• 2011

세계적 멸종위기종인 따오기(Nipponia nippon)는 2008년 10월에 중국에서 1쌍이 도입된 후 한국최초로 인공번식에 성공하였다. 본 연구는 따오기의 sex-related gene과 Chromodomain Helicase DNA Binding Protein gene (CHD gene)을 가지고 polymerase chain reaction (PCR)을 수행하여 새로 태어난 따오기 유조의 성별을 확인하고자 하였다. 본 연구에서는 따오기의 성별 확인을 위해 PCR후 제한효소의 처리 방법과 P2과 P8를 이용한 PCR 방법을 실시하였을 때 더 정확한 결과가 나타남을 알 수 있었다. 그리고 CHD gene의 염기서열을 선행연구와 비교해 본 결과, 암컷의 염기서열에서 1~2 base pairs 차이가 나타남을 알 수 있었다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.267-267
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• 1994

In order to obtain insight into the mechanism by which DNA containing a thymine photo-dimer is recognized by the excision repair enzyme, T$_4$ endonuclease V, we have taken NMR study of this protein and its complex with oligonucleotides. The conformations of five different DNA duplexes DNA I : d(GCGGATGGCG).d(CGCCTACCGC), DNA II d(GCGGTTGGCG) .d(CGCCAACCGC), DNA III : d(GCGGT ^ TGGCG) .d(CGCCAACCGC), DNA IV d(GCGGGCGGCG).d(CGCCCGCCGC) and DNA V d(GCGGCCGGCG) . d(CGCCGGCCGC) were studied by $^1$H NMR. The NMR spectra of these five DNA duplexes in the absence of the enzyme clearly show that the formation of a thymine dimer within the DNA induces only a minor distortion in the structure, and that the overall structure of B type DNA is retained. The photo-dimer formation is found to cause a large change in chemical shifts at the GC7 base pair, which is located at the 3'-side of the thymine dimer, accompanied by the major conformational change at the thymine dimer site. The binding of a mutant T$_4$ endonuclease V (E23Q), which is unable to digest DNA containing a thymine dimer, to the DNA duplex d(GCGGT ^ TGGCG)ㆍd(CGCCAACCGC) causes a large down-field shift in the imino proton resonance of GC7. Therefore, this position is thought to be either the crucial point of the interaction wi th T$_4$ endonuclease V, or the si to of a conformational change in the DNA caused by the binding of T$_4$ endonuclease V. Usually, it is very difficult to assign NMR peaks in DNA * protein complex because of severe peak overlaps. In order to overcome these peak overlaps, we used a method of deuterium incorporation.

• Clinical and Experimental Pediatrics
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• 제58권1호
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• pp.20-27
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• 2015

Purpose: We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. Methods: Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. Results: Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. Conclusion: We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.