• 한국어병학회지
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• 제5권2호
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• pp.143-152
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• 1992

Metallothionein은 세포내의 중금속의 농도을 조절하는 주요한 단백질로서 bacteria에서 척추동물에 이르기까지 모든 생명체에서 나타나는 공통된 단백질이다. 비록 metallothionein의 정확한 기능은 알려져 있지 않으나 독성을 나타내는 중금속에 대하여 세포내 방어기작에 관여할 뿐만 아니라 여러다른 유전자의 총괄적 조절기작 및 matalloprotein의 발현에 관여할 것으로 보고있다. 본 연구에서는 Channel Catfish의 metallothionein cDNA 유전자를 poly(A)를 갖는 mRNA로 부터 Reverse Transcriptase-Polymerase Chain Reaction(RT-PCR)에 의하여 cloning하였다. 증폭된 PCR products는 pBluescript SK+의 EcoRV site 및 pUC19의 Smal site에 dT tailing을 하여 cloning하였으며, PCR products는 multicloning site에 있는 EcoRI 및 HindIII 로 절단하여 확인하거나 신속한 PCR screening에 의하여 확인하였다. 여러 PCR clone 중 하나인 pMT150에 대한 DNA 염기서열을 조사한 결과 다른 어류의 metallothionein cDNA 유전자와 높은 유사성을 보였다.

• 대한바이러스학회지
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• 제27권2호
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• pp.185-195
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• 1997

Sequence comparison of the RNA-dependent RNA polymerase of human caliciviruses (HuCVs) from Korean children with gastroenteritis revealed significant genetic variation among them. cDNA clones were produced from the HuCVs collected from pediatric population during a period of 1987-1994. The application of reverse transcription-polymerase chain reaction (RT-PCR) using primers directed to the RNA-dependent RNA polymerase region within ORF1 of Norwalk virus (NV) showed that 13.7% of HuCVs yielded PCR products of similar size to the NV prototype, NV8FIIa/68/US, with exceptions of HuCV 185/87/Korea and HuCV 1115/90/Korea. Computer analyses showed that the PCR products had a continuous protein encoding frame on the positive strand, and contained GLPSG and YGDD amino acid motifs at the predicted distance from primers. Alignment of the amino acid sequences of HuCVs with previously published sequences for Snow Mountain agent (SMA), NV, and Sapporo/82/Japan indicated that these strains can be divided into four major genogroups. There were 10 (45%) SMA-like CVs, one (4.5%) NV-like HuCVs, two (9%) Sapporo-like HuCVs, and nine (41%) unidentified HuCVs. This fourth genogroup should be investigated further. HuCV 185/87/Korea and HuCV 1115/90/Korea, Sapporo-like CVs, were genetically distinct from previously characterized HuCVs and more closely related to known animal CVs. One of the animal CV-like strain, HuCV 185/87/Korea, showed nucleotide and amino acid homology of only 67% and 73% with the prototype Sapporo/82/Japan. Further characterization of animal and human CV genomes and studies of possible cross-transmission of CVs from animals to humans are likely to be beneficial in understanding the epidemiology of HuCVs.

• Journal of Plant Biotechnology
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• 제39권4호
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• pp.273-280
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• 2012

Differences of gene expression between red flash peach cultivar and white flash peach cultivar were investigated by Nest-generation sequencing (NGS). EST from the red flash peach cultivar and white flash peach cultivar were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, temperature stress, ethylene signal pathway were significantly higher in white flash peach cultivar than in red flash peach cultivar. On the other hand, the up-regulation of proteins involved in anthocyanin and flavonol biosynthesis and protein degradation and sorbitol metabolism were observed in red flash peach cultivar. Chalcone synthase was preferentially expressed in the red flesh peach cultivar, agreeing with the accumulation of anthocyanin and expression of other previously identified genes for anthocyanin biosynthesis. Anthocyanin pathway related genes CHS, F3H, DFR, LDOX, UFGT differentially expressed between red flash peach cultivar and white flash peach cultivar. These results suggest that red flash peach cultivar and white flash peach cultivar have different anthocyanin biosynthesis regulatory mechanisms.

• 한국동물위생학회지
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• 제37권3호
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• pp.219-224
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• 2014

A eight-month-old blue and yellow macaw (Ara ararauna) with psittacine beak and feather disease (PBFD)-suspected signs, such as, abnormal feather, depression and diarrhea, was presented to Animal Disease Intervention Center, Kyungpook National University in 16 April 2014. The partial ORF V1 gene of PBFD virus (PBFDV) was detected by polymerase chain reaction (PCR) from DNA templates extracted from feather, blood and cloacal swab sample of the bird, but no other viral DNAs that often infected in psittacine birds including avian bornavirus and avian polyomavirus were detected from the samples of the bird, indicating this case is due to single infection of PBFDV. Nucleotide sequence analysis of the amplified partial ORF V1 gene was confirmed to have 96.7% and 93.6% homology with that of previously reported PBFDV strain (Genbank no. HM748924 and FJ685980). This report describes the first detection of PBFDV in PBFD-suspected blue and yellow macaw in Korea.

• 한국동물위생학회지
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• 제37권3호
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• pp.213-218
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• 2014

In early April 2014, a month-old Alexandrine Paraqeet (Psittacula eupatria) that was raised in a domestic aviary located in Gyungju-si, Korea was suddenly died and submitted to Animal Disease Intervention Center, Kyungpook National University in order to diagnose the causative agent. In post-mortem examination, the bird had abnormally developed feathers on the neck and abdomen region and subcutaneous hemorrhages on the neck and cheek adjacent to the beak. At necropsy, the bird had hemorrhage on the muscle of the femoral region, ascites, multi-focal hemorrhages on the epicardium, and diffuse hemorrhages on the sub-serosa of proventriculus and gizzard, suggesting typical avian polyomavirus (APV) infection. The partial large tumor (T) antigen gene of APV was detected by PCR from tissues of the heart, lung, liver, kidney, proventriculus and feathers of the APV-suspected birds. However, other pathogenic virus-specific nucleic acid common with psittacine birds such as avian bornavirus, psittacine beak and feather disease virus and psittacid herpesvirus were not detected from the mixed tissue samples of the bird, indicating this case is due to single infection of APV. Nucleotide sequence analysis of the partially amplified large T antigen DNA was confirmed to have 99~100% homology with that of the previously reported APV strains. This case report describes the first detection of APV in Alexandrine Paraqeet in Korea.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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• pp.63-67
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• 1999

We have isolated more than a dozen cDNA clones corresponding to genes that were expressed in Arabidopsis leaves when they were kept in the dark. The nucleotide sequence analysis showed that some of the clones encoded proteins with significant homology to $\beta$-glucosidase (din2), branched-chain $\alpha$-keto acid dehydrogenase subunit E1$\beta$(din3), and another subunit E2 (din4), yeast RAD23 (din5), asparagine synthetase (din6), pre-mRNA splicing factor SRp35 (din7), phosphomannose isomerase (din9), seed imbibition protein (din10), and 2-oxoacid-dependent oxidase (din11). Accumulation of transcripts from din3,4,6 and 10 occurred rapidly after the plants were transferred to darkness. Transcripts from din2,9, and 11 could be detected only after 24 h of dark treatment. Inhibition of photo-synthesis by DCMU strongly induced the accumulation of transcripts from those genes, and application of sucrose to detached leaves suppressed the accumulation both in the dark and by DCMU. These observations indicate that expression of the genes is caused by sugar starvation resulted from the cessation of photosynthesis. We further showed that din2-encoded protein also accumulated in senescing leaves. Given these results, possible roles of din genes in leaves in the dark and senescing leaves are discussed.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.274-282
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• 2019

Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

• 한국동물위생학회지
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• 제45권3호
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• pp.211-219
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• 2022

Bovine viral diarrhea (BVD) is one of the problematic wasting diseases in cattle leading to huge economic losses. This study was conducted to investigate the prevalence of BVD including transient and persistent infection from cattle farms in Gyeongsangnam-do. A total of 2,667 blood samples from 24 farms were collected and the sera were subjected to ELISA to detect BVD virus (BVDV) antigen, E^rns. 5' untranslated region (5'-UTR) of BVDV-positive samples was sequenced to identify the genotype, and compared with isolates previously reported elsewhere. There were fourteen BVDV-positive calves from 2,667 samples (positive rate: 0.52%) from first ELISA testing followed by eight persistently infected out of eleven BVDV-positive samples (72.73%) in secondary ELISA that was conducted in at least four weeks suggesting the circulation of BVDV in the area. Sequencing analysis exhibited that thirteen BVDV-positive samples were identified as BVDV-1b and one sample was BVDV-2a. Phylogenetic analysis revealed that the BVDV-1b-positive samples showed the highest homology in nucleotide sequence to Korean isolates collected from Sancheong, Gyeongsangnam-do, while the BVDV-2a-positive sample (21GN7) was more similar to reference strains collected outside South Korea. This study will provide the recent fundamental data on BVD prevalence in Gyeongsangnam-do to be referred in developing strategies to prevent BVDV in South Korea.

• 한국미생물·생명공학회지
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• 제32권4호
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• pp.297-306
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• 2004

열 충격단백질(Heat shock protein : HSP)은 온도 스트레스에 대하여 세포 내에서 발현되는 단백질이다. HSP의 중요 분류군의 하나가 HSP90 family 이다. 여러 종류의 포유동물과 조류에서 HSP 유전자 특성에 대한 연구가 많이 진행되었다. 본 연구에서는 넙치(Paralichthys olivaceus)로부터 제조한 넙치 뇌 cDNA 유전자 은행을 이용하여 넙치 HSP90 cDNA 유전자를 분리하여 구성 염기서열의 특성을 밝혀 내었다. 염기서열의 분석결과 넙치의 hsp90$\beta$ 유전자는 2,791 개의 뉴클레오타이드로 구성되어 있고, 726개의 아미노산 잔기가 암호화되어 있었다. 넙치 hsp90$\beta$ 유전자는 European sea bass와 96.6% zebrafish와 92.9%, Atlantic salmon와 92.0%, 그리고 사람과는 89.5%의 염기서열 상동성을 지니고 있었다. 또한 HSP90 아미노산 서열을 바탕으로 척추동물 종들과의 진화계통수를 구축하였다. 넙치 hsp90$\beta$ 유전자의 mRNA의 분포 정도를 RT-PCR를 이용하여 조사하였다. hsp90$\beta$ 유전자는 조사한 모든 조직(뇌, 간, 신장, 근육, 비장)에서 높은 수준으로 발현이 되고 있었다. 또한, 넙치 hsp90$\beta$ 단백질을 대량발현하기 위하여 대장균에서 발현을 유도하였다.

• 농업과학연구
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• 제36권2호
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• pp.167-177
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• 2009

본 연구는 한국형 Holstein종 젖소집단의 DGAT1 유전자의 특성을 구명하고, DGAT1의 유전적 다형과 산유형질인 유량 및 유지량 간의 연관성을 구명하여 젖소집단의 유전적 개량을 위한 분자유전학적 접목을 위하여 실험을 실시하였다. Holstein종의 genomic DNA로부터 PCR기법을 이용하여 DGAT1 유전자좌를 specific primers로 증폭한 후, 1.5% agarose gel에 전기영동한 결과 411 bp의 단편이 양호하게 증폭되었음을 확인하였다. DGAT1 유전자의 염기서열을 분석한 결과 DGAT1 Q 대립유전자의 216-218 bp의 염기서열이 AUG(lysine, K)였으나, 동위치의 대립유전자 q의 염기서열은 GCG(alanine, A)로 치환되어 있음을 확인할 수 있었다. 한편 한국 Holstein종과 NCBI에서 보고된 bovine DGAT1 유전자 단편의 염기서열간에는 100% 상동성을 보였다. 한국 Holstein종 유우집단의 DGAT1 유전자형의 분포는 DGAT1 QQ, Qq 및 qq 유전자 분포가 각각 16.43, 36.43 및 47.14%로 qq 유전자형빈도가 다른 유전자형에 비하여 높았으며, 유전자빈도는 DGAT1 Q 및 q 빈도가 각각 0.35 및 0.65로 q의 빈도가 높았다. DGAT1 유전자형과 산유량인 유량 및 유지량간의 연관성에 있어서는 DGAT1 유전자형이 유량 및 유지량에서 유의적인 차이 (P<0.05)를 보였으며, DGAT1 Qq 유전자형이 QQ 및 qq 유전자형에 비하여 유량과 유지량에서 유의적인 차이(P<0.05)로 높은 수치를 나타냈다.