• 식물분류학회지
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• 제43권1호
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• pp.12-21
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• 2013

본 연구에서는 제주황기(Astragalus nakaianus)의 분류학적 위치를 명확히 하고 한국산 재배황기(A. mongholicus cultivar)의 올바른 학명을 부여하기 위하여 외부형태형질과 ITS 그리고 5구간의 cp non-coding DNA의 염기서열을 조사하였다. 또한 9개의 마이크로새털라이트 마커를 이용하여 3집단 61개체에 대한 유전적 구조가 분석되었다. 그 결과, 남한산 재배황기와 A. mongholicus var. dahuricus 사이에서는 형태와 ITS 염기서열상에서 유의한 차이점이 없었다. 제주황기는 A. mongholicus var. mongholicus 그리고 var. dahuricus와 줄기의 습성, 식물체의 길이, 엽축의 길이, 소엽의 길이 등에서 형태적으로 차이를 보였다. ITS와 cp non-coding 구간 염기서열에서 제주황기는 황기(A. mongholicus)와 차이점을 보이지 않았지만, 마이크로새털라이트 분석에서 제주황기와 남한산 재배황기간에 뚜렷하게 구분된 구조를 보였다. 이러한 결과들을 보았을 때, 한국산 재배황기는 A. mongholicus var. dahuricus로 처리해야 하며 제주황기는 A. mongholicus의 변종으로서 취급되어야 한다.

• 농업과학연구
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• 제20권2호
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• pp.189-200
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• 1993

Ornithine Decarboxylase는 동물세포에 있어서 polyamines을 생합성하는 반응의 율속 단계에 있는 효소이다. 세포의 성장기간 동안에 이 효소는 mRNA와 단백질 수준에서 급격한 변화를 일으키며 조절된다. 이와 같은 polyamines농도의 급변화를 분자수준에서 탐구하기 위하여 ODC 특이 cDNA colne이 cDNA libray에서 분리되었다. cDNA의 clone이 이루어진 부분은 open reading frame과 3'-noncording region, 그리고 poly A의 tail이 있는 부분으로서 각각 456, 348, 14개의 nucleotides로서 구성되어 있었다. 인간, mouse, rat, hamster로부터 같은 지역내 추론된 C-말단으로부터의 아미노산의 비교는 단백질 상동성에 88%이상을 보였다. 이와같은 아미노산 염기순서에 있어서의 잘 보존된 특징은 ODC가 세포성정과 분화에 있어서 중요한 역할이 있음을 잘 나타내고 있다.

• Animal cells and systems
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• 제3권4호
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• pp.393-397
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• 1999

Length variations in human mitochondrial DNA (mtDNA) offer useful markers in the study of female aspects of human population history. One such length variation is a 9-bp deletion in the small noncoding segment located between the COII and Iysine tRNA genes (COII/tRNA/$^{Lys}$ intergenic region) which usually contain two tandemly arranged copies of a 9-bp sequence (ccccctcta) in human mtDNA. The mtDNA 9-bp deletion and polymorphic variants of expanded 9-bp repeat motif in the intergenic COII/tRNA$^{Lys}$ region have been found at varying frequencies among different human ethnic groups. We have examined the length variation of the mtDNA COII/tRNA$^{Lys}$ intergenic region from a total of 813 individuals in east Asian populations. The occurrence of the 9-bp deletion was found to be relatively homogeneous in northeast Asian populations (Chinese, 14.2%; Japanese, 14.3%: Koreans, 15.5%), with the exception of Mongolians (5.1%). In contrast, Indonesians (25.0%) and Vietnamese (23.2%) of the southeast Asian populations appeared to have relatively high frequencies of the 9-bp deletion. We identified the existence of a new expanded 9-bp repeat motif which likely resulted from a slipped mispairing insertion of six more cytosines in the intergenic COII$^{Lys}$ region. It was present at low frequencies in the Korean (2/349) and Japanese populations (2/147). Based on the results of this study, the Korean population may reflect a close genetic affinity with the Japanese and Chinese populations than the others surveyed east Asian populations.

• 한국미생물·생명공학회지
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• 제13권1호
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• pp.19-25
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• 1985

Saccharomyces cerevisiae HIS 5 유전자는 histidinol phosphate aminotransferase (EC: 2, 6, 1,9)를 code하는 아미노산 합성유전자이다. 이 유전자는 plasmid pSH 530에 cloning되어 E. coli와 Saccharomyces cerevisiae 숙주에서 promoter로서 전사하였다. HIS 5 유전자의 총염기 수는 736개이였고 5' 상류영역에는 긴 reading frame, directed repeat, 전사개시점, 그리고 Pribnow box염기배열이 있었다. 특히 HIS 5 유전자의 ATG 주변 염기배열은 -A-A-A-T-T-A-C-A-C-T-A-T-G-G-T-T-T-T-T-G-A-T-였으며 C block은 없었다.

• 대한바이러스학회지
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• 제26권2호
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• pp.215-225
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• 1996

Aseptic meningits, an acute inflammation of the meninges, is a common illness during childhood. Virus is the most important cause of aseptic meningitis. Especially enterovirus causes approximately above 85% of all cases of aseptic meningitis. In 1993, there was a big epidemic of aseptic meningitis by ECHO 9 and ECHO 30 viruses. And ECHO 3 virus was isolated as a causative agent of aseptic meningitis in 1994. This study was aimed to detect the causative agent of aseptic meningitis in 1995 and to analyze the 5'-noncoding region which was used to detect virus. Virus was isolated from 87 stools and cerebrospinal fluid specimens of the patients by cultured RD and HEp-2 cell. Neutralizing antibody tests using enterovirus serum pool were performed on the specimens with cytopathic effect. 3 of ECHO 7 viruses and 5 of Coxsackie B3 viruses were isolated from stool specimens and 1 of ECHO 7 and Coxsackie B3 mixed type was confirmed from cerebrospinal fluid specimens. RNA was isolated from the culture supernatants of infected cells and general primers were selected in highly conserved part of the 5'-noncoding region of the enteroviral genome for RT-PCR. PCR product from this virus showed a 152bp band on gel electrophoresis. Sequence of obtained DNA was compared with prototype sequences by accessing to the Genebank database. 5'-noncoding region of isolated Coxsackie B3 virus, which has point mutations in nucleotide sequence positions 493, 497, 502, 523, was closely related to that of polio virus type 1, Mahoney strain. In case of isolated ECHO 7 virus, nucleotide has been changed from cytosine to thymine at position 581 and from thymine to cytosine at position 583. We concluded the causative agents of the outbreak of aseptic meningitis during June to July in 1995 were both ECHO 7 and Coxsackie B3 virus, and the primer used in this study could allow a rapid diagnosis of enteroviruses by PCR.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1800-1807
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• 2016

To understand how human cytomegalovirus (HCMV) might change and evolve after reactivation, it is very important to understand how the nucleotide sequence of cultured HCMV changes after in vitro passaging in cell culture, and how these changes affect the genome of HCMV and the consequent variation in amino acid sequence. Strain JHC of HCMV was propagated in vitro for more than 40 passages and its biological and genetic changes were monitored. For each passage, real-time PCR was performed in order to determine the genome copy number, and a plaque assay was employed to get virus infection titers. The infectious virus titers gradually increased with passaging in cell culture, whereas the number of virus genome copies remained relatively unchanged. A linear correlation was observed between the passage number and the log₁₀ infectious virus titer per virus genome copy number. To understand the genetic basis underlying the increase in HCMV infectivity with increasing passage, the whole-genome DNA sequence of the high-passage strain was determined and compared with the genome sequence of the low-passage strain. Out of 100 mutations found in the high-passage strain, only two were located in an open reading frame. A G-T substitution in the RL13 gene resulted in a nonsense mutation and caused an early stop. A G-A substitution in the UL122 gene generated an S-F nonsynonymous mutation. The mutations in the RL13 and UL122 genes might be related to the increase in virus infectivity, although the role of the mutations found in noncoding regions could not be excluded.

• 미생물학회지
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• 제32권2호
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• pp.120-125
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• 1994

Vibrio meschnikovii 균주 RH530의 trpB. trpA 및 3‘ trpC(F) 유전자를 대장균에 클로닝하여 염기서열을 결정하였다. trpB 및 trpA 유전자는 각각 391 및 268 아미노산을 coding할 수 있는 1,173 bp 및 804 bp의 open reading frame을 가졌다. trpB 및 trpA 유전자는 일반적인 ribosome-binding sequence를 가졌으며 1개의 nucleotide만큼 중복되어 있었다. 이는 이들 두 유전자가 trinslation 단계에서 연결되어 있음을 의미한다. trpB 유전자의 개시토돈에서 115 nucleotide 위에 trpC(F)와 trpF의 융합체인 trpC(F)의 3’-말단부위의 불완전한 open reading frame의 존재 하였다. V. metschnikovil RH530의 TrpB, TrpA 및 TrpC(F)의 아미노산 서열은 V. parahaemolyticus의 그것들과 각각 64.2%, 82.4%, 73.7%: S. typhimurium과 58.7%, 72.3%, 54.9%: B. lactoermentum과 42.6%, 54.1%, 12.5%의 유사성을 보였다. 이는 TrpB가 TrpA보다 서열이 잘 보존되어 있음을 나타내며 TrpC(F)는 다를 두 polypeptide에 비해서 서열의 변이가 큼을 나타낸다. 이들 유전자들의 구조, 특히 trpC(F)와 trpB 사이의 noncoding 부위는 Enterobacteriaceae 비롯한 다를 개체들과는 다른 특징을 보였다.

• Applied Biological Chemistry
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• 제38권1호
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• pp.55-62
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• 1995

한국 마늘 바이러스의 유전자 구조와 병 발생 메카니즘을 연구하기 위하여, 바이러스가 감염된 마늘잎으로부터 바이러스 입자를 분리하고 RNA를 추출하였다. 그 virus RNA를 이용하여 마늘 바이러스 cDNA 유전자 은행을 만들어 일부 clone의 염기 서열을 결정하였다. 여기에서 얻은 cDNA clones 중에서 poly(A) tail을 갖는 clone S81를 분리하고 873 bp의 전체 염기서열을 결정하였다. Clone S81의 염기서열을 다른 식물 바이러스와 비교한 결과 potexvirus의 껍질단백질 부분의 염기서열과 $30{\sim}40%$의 유사성을 보여주었다. Clone S81은 바이러스 RNA의 3' 말단 부위에 해당하고, 껍질단백질의 N-terminal 3개 아미노산이 빠진 open reading frame (ORF) 및 3'-noncoding region을 포함하고 있다. 3' 말단 부분에는 바이러스 복제과정에서 cis-acting element로 작용한다고 여겨지는 hexamer motif와 polyadenylation signal이 존재한다. 이 clone을 probe로 하여 Northern blot을 실시한 결과 genome의 크기는 7.5 knt라는 것을 알 수 있었고 clone S81은 potexvirus의 cDNA clone이라는 결론을 얻었다. 한국 마늘에서 이 바이러스의 분포 양상을 알아보기 위해 껍질단백질에 대한 항체를 만들었다. 먼저 발현벡터를 이용하여 대장균에서 대량으로 발현시키고 affinity chromatography로 껍질단백질을 정제하였다. 그 단백질을 토끼에 주사하여 껍질단백질에 대한 항체를 얻었다. 이 항체를 사용하여 다양한 지역에서 재배되는 마늘잎의 추출액에 대해 immunoblot을 실시하였다. 그 결과 분자량 29,000과 27,000 위치에서 signal을 보였다. 분자량 27,000 단백질이 29,000이 분해되어 생긴 산물인지 알아보기 위하여 그 추출액을 $37^{\circ}C$에서 시간을 달리하여 incubation한 후 immunoblotting하였다. 그 결과도 마찬가지로 같은 위치에서 signal을 보여줬다. 따라서 한국 마늘에는 재배되는 지역에 따라 다소 다르기는 하지만 대체로 두 종류의 potexvirus로 감염되어 있다고 추정된다.

• Journal of Microbiology
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• 제44권1호
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• pp.54-63
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• 2006

Ferritin is a major eukaryotic protein and in humans is the protein of iron storage. A partial gene fragment of ferritin (255 bp) taken from the total RNA of Periserrula leucophryna, was amplified by RT-PCR using oligonucleotide primers designed from the conserved metal binding domain of eukaryotic ferritin and confirmed by DNA sequencing. Using the $^{32}P-labeled$ partial ferritin cDNA fragment, 28 different clones were obtained by the screening of the P. leucophryna cDNA library prepared in the Uni-ZAP XR vector, sequenced and characterized. The longest clone was named the PLF (Periserrula leucophryna ferritin) gene and the nucleotide and amino acid sequences of this novel gene were deposited in the GenBank databases with accession numbers DQ207752 and ABA55730, respectively. The entire cDNA of PLF clone was 1109 bp (CDS: 129-653), including a coding nucleotide sequence of 525 bp, a 5' -untranslated region of 128 bp, and a 3'-noncoding region of 456 bp. The 5'-UTR contains a putative iron responsive element (IRE) sequence. Ferritin has an open reading frame encoding a polypeptide of 174 amino acids including a hydrophobic signal peptide of 17 amino acids. The predicted molecular weights of the immature and mature ferritin were calculated to be 20.3 kDa and 18.2 kDa, respectively. The region encoding the mature ferritin was subcloned into the pT7-7 expression vector after PCR amplification using the designed primers and included the initiation and termination codons; the recombinant clones were expressed in E. coli BL21(DE3) or E. coli BL21(DE3)pLysE. SDS-PAGE and western blot analysis showed that a ferritin of approximately 18 kDa (mature form) was produced and that by iron staining in native PAGE, it is likely that the recombinant ferritin is correctly folded and assembled into a homopolymer composed of a single subunit.

• International Journal of Industrial Entomology and Biomaterials
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• 제44권2호
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• pp.55-64
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• 2022

Bombyx shini Park & Sohn, 2002 (Lepidoptera: Bombycidae), which was listed as an endemic species in South Korea has recently been renamed as the East Asian silk moth Rotunda rotundapex Miyata & Kishida, 1990 (Lepidoptera: Bombycidae). In this study, we sequenced the complete mitochondrial genome (mitogenome) of the R. rotundapex to announce genomic characteristics and to clarify its validity with a new name. The 15,294-bp long complete mitogenome comprises a typical set of genes [13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes] and one major noncoding, A + T-rich region, with an arrangement identical to that observed in most lepidopteran mitogenomes. The A/T content of the whole mitogenome was 79.22%; however, it varied among the regions/genes as follows: A + T-rich region, 91.62%; srRNA, 84.67%; lrRNA, 83.01%; tRNAs, 81.43%; and PCGs, 77.46%. Phylogenetic analyses of 35 species in the Bombycoidea superfamily showed the sister relationship between the families Sphingidae and Bombycidae s. str., with the higher nodal support [bootstrap support (BS) = 78%]. The Saturniidae was placed as the sister to the two families, but the nodal support for this relationship was low (BS = 53%). Current R. rotundapex was placed together with previously reported con-species with the highest nodal support, forming a separate clade from Bombyx, validating that B. shini can have a new genus name, Rotunda. However, the Korean R. rotundapex showed a substantial sequence divergence at 5.28% to that originated from an individual of type locality Taiwan in 1,459-bp of COI sequences. Considering such a high sequence divergence an additional study, which includes morphological and DNA barcoding data from further extensive distributional range maybe is needed for further robust taxonomic conclusion.