• Reproductive and Developmental Biology
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• 제34권4호
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• pp.283-288
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• 2010

During normal early embryonic development in mammals, the global pattern of genomic DNA methylation undergoes marked. changes. The level of methylation is high in male and female gametes. Thus, we cloned the cDNA of the porcine DNA methyltransferase 1 (Dnmt1) gene to promote the efficiency of the generation of porcine clones. In this study, porcine Dnmt1 cDNA was sequenced, and Dnmt1 mRNA expression was detected by reverse transcription-polymerase reaction (RT-PCR) in porcine tissues during embryonic development. The porcine Dnmt1 cDNA sequence showed more homology with that of bovine than human, mouse, and rat. The complete sequence of porcine Dnmt1 cDNA was 4,774-bp long and consisted of an open reading frame encoding a protein of 1611 amino acids. The amino acid sequence of porcine DNMT1 showed significant homology with those of bovine (91%), human (88%), rat (76%), and mouse (75%) Dnmt1. The expression of porcine Dnmt1 mRNA was detected during porcine embryogenesis. The mRNA was detected at stages of porcine preimplantation development (1-cell, 2-cell, 4-cell, 8-cell, morula, and blastocyst stages). It was also abundantly expressed in tissues (lung, ovary, kidney and somatic cells). Further investigations are necessary to understand the complex links between methyltransferase 1 and the transcriptional activity in cloned porcine tissues.

• 미생물학회지
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• 제30권5호
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• pp.397-402
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• 1992

서울지역의 소아설사환자가 가검물로부터 분리한 로타바이러스의 VP7을 코딩하는 RNA분절 cDNA를 합성한 후 로타바이러스 혈청형1인 WA1과 RE9의 아홉 번째 RNA분절과 비교하였더니 90%이상의 유사성을 보였다. 염기서열로부터 유추된 아미노산 서열중 혈청간에 변이가 많은 VR5와 VR8 지역을 비교한 결과 역시 혈청형 1인 RE9과 WA1 바이러스주와 매우 높은 유사성을 지님을 알 수 있었다.

• Applied Biological Chemistry
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• 제32권1호
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• pp.64-72
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• 1989

Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

• 한국정보과학회:학술대회논문집
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• 한국정보과학회 2003년도 가을 학술발표논문집 Vol.30 No.2 (2)
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• pp.772-774
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• 2003

DNA/DNA의 연쇄 결합 반응에 대한 시뮬레이션을 열역학적 데이터를 이용하여 구현하였다. 1-Base의 non Watson-Crick 결합과, dangling end(결합이 이루어진 두개의 DNA strand 중 한쪽 끝이 다른 쪽 끝보다 길거나 짧은 경우)를 허용하는 nearest-neighbor model을 사용하여 구현된 이 모델에서는 한번의 hybridization만을 예측하는 것이 아니라 연속적인 결합 반응의 시뮬레이션이 가능하다. 이를 통해서 분자 알고리즘의 설계와 검증이 가능할 뿐만 아니라, cross-homology의 검사를 통한 시퀀스의 검증까지도 가능하다. 이러한 in silico 에서의 접근 방식은 효율적인 분자 알고리즘의 개발과 신뢰성 있는 시퀀스의 설계에 도움이 될 수 있다.

• Pediatric Infection and Vaccine
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• 제6권1호
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• pp.123-130
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• 1999

목 적 : 무균성뇌막염은 소아에서 주로 발생하는 질환으로, 흔히 장 바이러스에 의해 초래 된다. 저자들은 그동안 우리나라에서 분리된 무균성뇌막염 원인바이러스의 5'-NCR에 대한 sequencing을 통하여 prototype과의 homology를 비교하고 이 연구를 진단에 이용하기 위한 기초자료로 이용하기 위하여 본 연구를 시도하였다. 방 법 : 과거 4년간 우리나라에서 분리한 장바이러스 Coxsackie B1, Echovirus 3, 7, 9, 30 을 이용하여 RNA를 분리하고 RT-PCR을 이용하여 DNA를 합성한 후, direct sequencing을 이용하여 WHO에서 얻은 prototype과 homology를 비교하였다. 결 과 : 1) PCR product는 155bp와 440bp부위에 특징적인 띠를 관찰할 수 있었다. 2) 155bp에 관한 sequencing homology를 보면 prototype의 Coxsackie virus와 echovirus 는 92.1%의 homology를 보였다. 3) 환자에서 분리한 Coxsackie B1은 prototype과 94.1%의 homology를 보였다. 4) 환자에서 분리한 Echovirus 3은 92.8%, Echovirus 7은 92.8%, Echovirus 9는 94.1%, Echovirus 30은 82.9%의 homology를 보였다. 결 론 : 장바이러스의 5'-NCR은 homology가 높아서 진단에 이용하기에 좋으며 이 부위를 통한 typing을 위해서는 더 긴 부위를 sequencing할 필요가 있다.

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.63-68
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• 2002

A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

• 한국수산과학회지
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• 제36권1호
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• pp.35-38
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• 2003

Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya tenera (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1,822 bp exon and a 510 bp intron. The G+C contents of exon and intron were $48.68\%\;and\;54,90\%,$ respectively. The exon sequence showed $99.6\%$ homology to the GebBank accession number AB029880 of the Japanese P. tenera. The intron region that is inserted upstream between 568 and 1,079 showed $43.6\%$ homology to the AB029880.

• The Plant Pathology Journal
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• 제19권2호
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• pp.106-110
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• 2003

Stolbur Phytoplasma was detected from Lilium Oriental hybrids showing flattened stem and flower clustering. The presence of phytoplasma was demonstrated using polymerase chain reaction（PCR) assays with phyto-plasma-universal（P1/P6）and stolbur phytoplasma-specific 16F1/R1-S primer pairs amplifying phytoplasma 16S rDNA regions. Nucleotide suquences of the phytoplasma 16S rDNA were determined. Nucleic acid extracted from lily amplified 1.5 kb DNA with a phytoplasma universal primer pair. In nested PCR, 1.1 kb PCR product was obtained using specific primer pair, indicating an isolate of stolbur phytoplasma. Nucleotide sequence of phytoplasma 16S rDNA reported in this study showed 99.5% and 99.1％ identities with two known stolbur phytoplamas (16Sr XII-A). Also, it exhibited a sequence homology of 98.0％ with phormium yellow leaf (16Sr XII-B), and 97.9％ with Australian grapevine yellows (16Sr XII-B). Meanwhile, it showed 98.1% identity with strawberry green petal phytoplama, (16Sr1-C), and 94.7 % with American aster yellows (16Sr1-B). Homology percentage of the 16S rDNA nucleotide sequence suggests that this phytoplama could be classified into the stolbur phytoplasma, subgroup A (16Sr XII-A), as a type strain stolbur.

• 식물조직배양학회지
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• 제26권1호
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• pp.41-47
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• 1999

토마토의 genomic DNA library로부터 분리한 tPALl, tPAL4유전자의 염기서열을 분석하여 tPAL5 유전자와 비교 분석한 결과는 다음과 같다. tPAL5 유전자는 722개의 아미노산과 710 bp의 intron을 가지고 있으나 tPALl은 intron을 가지고 있지 않으며 또한 tPAL5 유전자와 비교하여 249개의 짧은 polypeptide를 가지고 있었다. tPAL4유전자인 경우 357개의 아미노산과 305bp의 intron을 가지고 있었다. tPAL 효소간의 아미노산 homology는 tPAL1유전자와 tPAL4 유전자간은 87.2%, tPALl과 tPAL5는 85.3%, tPAL4 와 tPAL5 는 91.4%의 homology를 보였다. 또한, tPALl, tPAL4 유전자는 정상적인 polypeptide를 가지는 tPAL5유전자와 비교하여 비정상적인 stop codon을 가진 짧은 polypeptide로 구성되어 있었다. 다양한 식물 종으로부터 분리된 PAL유전자의 염기서열을 비교한 결과 토마토 (Lycopersicon esculentum), 감자 (Solanum tuberosum), 고구마 (Ipomoea batatas)간의 유연관계과 높았으며, parsley (Petroselinum crispum), bean (Phaseolus vulgaris), pea (Pisum sativum), alfalfa (Medicago sativa) 등이 각각 서로간에 유연관계가 높았다. 또한, 토마토에서 분리한 family내에서 tPAL4와 tPAL5 유전자는 homology가 매우 높았고 (93.0%), tPAL1와 tPAL4유전자 사이는 다소 낮았으며 (84.4%), 특히 tPAL4는 감자의 PAL 유전자와 매우 높았다 (90.6%).

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.369-377
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• 1998

Rotavirus is a major cause of severe gastroenteritis in young children and animals throughout the world. The VP7 of rotavirus is thought to induce the synthesis of neutralizing antibodies and to be responsible for determining viral serotypes. The cDNA coding for the VP7 capsid protein of human rotavirus, obtained from Korean patients (HRV-Y14), was cloned and its nucleotide sequence was determined. Comparative analysis of the nucleotide sequences between VP7 of Y14 and that of other foreign isolates showed $92.7~95.2\%$ homology to G1 serotypes (RV-4, KU, K8, WA), $74.2\%$ homolgy to G2 serotype HU-5, $76.4\%$ homology to G3 serotype SA-11, and $77.6\%$ homology to G4 serotype A01321. These data suggest that HRV-Y14 can be classified as a G1 serotype. cDNA coding for VP7 of HRV-YI4 was subcloned into the baculovirus vector and the VP7 glycoprotein was expressed in insect cells. The expressed proteins in Sf9 cell extract and tissue culture fluid were separated on SDS-PAGE, and Western blot analysis with monoclonal antibody raised against the synthetic peptide containing 21 amino acids within the VP7 conserved region was performed. The molecular weight of recombinant VP7 was estimated to be 36 kDa which is about the same size as the native VP7. Addition of tunicamycin in the culture media caused a reduction of the molecular weight of the recombinant VP7 indicating that the expressed protein was glycosylated.