• The Plant Pathology Journal
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• 제18권4호
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• pp.187-191
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• 2002

This paper describes the cloning and sequence analysis of the 5'-terminal region and full-length cDNA production of genomic RNA of Lily symptomless virus (LSV), a Species Of the genus Carlavirus. A sing1e DNA band about 600 bp harboring the 5'-end of genomic RNA of the virus was successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), and was cloned for nucleotide sequence determination. Sequence analysis of selected RACE cDNA clones revealed that the LSV 5'non-translated region consists of 67 nucleotides long of AT rich stretch followed GC rich from the 5'-end. To produce full-length cDNA products for the viral genomic RNA, a set of LSV-specific primers could be designed based on the obtained sequence in this study and the known sequences of 3'-terminal region for the virus. Full-length cDNA copies of LSV, an 8.4 kb long, were directly amplified by the long-template RT-PCR technique from the purified viral genomic RNA samples. This full-length cDNA copies were analyzed by restriction mapping. The molecules produced in this study can be useful for the production of in vitro infectious cDNA clone, as well as, for the completion of genomic RNA sequence and genome structure for the virus.

• International Journal of Industrial Entomology and Biomaterials
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• 제4권2호
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• pp.163-168
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• 2002

We report for the first time the cDNA sequence encoding a ferritin subunit from the spiders Araneus ventricosus. The complete cDNA sequence of A. ventricosus ferritin subunit comprised 516 bp with 172 amino acid residues. The A. ventricosus ferritin subunit cDNA contained a conserved iron responsive element sequence in the 5 untranslated region. An alignment of the deduced protein sequence of the A. ventricosus ferritin subunit gene to that of other heavy chain ferritin molecules showed that A. ventricosus ferritin subunit is most similar to the great pond snail, Lymnaea stagnalis, ferritin with 70.2% of protein sequence identity.

• 한국식물병리학회지
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• 제14권3호
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• pp.240-246
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• 1998

A cDNA encoding desiccation-related protein was isolated from a flower bud cDNA library of Chinese cabbage (C-DH) and its nucleotide sequence was characterized. It contains 679 bp nucleotides with 501 bp open reading frame. The amino acid sequence of the putative protein showed the highest amino acid sequence homology (79 % identity) to dehydrin protein in Gossypium hirsutum. Also, the C-DH shares 48-52% amino acid sequence identity with the other typical dehydrin proteins in plant cells. When the amino acid sequence of their proteins were aligned, several peptide motifs were well conserved, of which function has to be solved. Particularly the C-DH contains 15 additional amino acids at its N-terminus. Genomic Southern blot analysis using the coding region of C-DH showed that the C-DH consists of a single copy gene in Chinese cabbage genome. The C-DH mRNA, whose transcript size is 0.7 kb, was expressed with a tissue-specific manner. It was highly expressed in seed, flower buds and low expression as detected in root, stem or leaf tissues of Chinese cabbage. And the transcript level of C-DH was significantly induced by the treatment of plant hormone, abscisic acid and water-deficit conditions.

• Journal of Plant Biotechnology
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• 제2권3호
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• pp.115-121
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• 2000

A cDNA clone encoding chloroplast triosephosphate isomerase (TPI-cp) was isolated from strawberry fruit cDNA library. Sequence analyses indicated that the cDNA contains an open reading frame of 314 amino acids (33.5 kDa) composed of a transit peptide (59 amino acids) in amino terminal region and mature protein (255 amino acids). The existence of transit peptide in the deduced amino acid sequence implies that it encodes a chloroplast isoform. The protein sequence is more similar to other plant chloroplast isoforms than cytosolic isoforms. RNA blot analysis indicated that its expression is ubiquitous in examined five tissues, flowers, leaves, petioles, roots and fruits, and shows differential pattern according to fruit ripening. Genomic DNA blot analysis showed that TPI-cp is encoded by multiple genes in strawberry. Through sequence comparison and phylogenetic tree construction, TPI-cp is distinctively grouped into dicot and chloroplast isoforms.

• 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.

• BMB Reports
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• 제32권5호
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• pp.502-505
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• 1999

cDNA fragments of ovine liver pyridoxal kinase were amplified by PCR using degenerate oligonucleotide primers derived from partial amino acids sequences of the enzyme. Using PCR products as probes, several overlapping cDNA clones were isolated independently from an ovine liver and a human brain cDNA library. The largest cDNA clone for each was selected for sequence analysis. The ovine liver cDNA encodes a polypeptide of 297 amino acid residues with Mr of 32,925, whereas the human clone is comprised of an open reading frame encoding 312 amino acid residues with Mr of 35,102. The deduced sequence of the human brain enzyme is completely identical to that of human testes cDNA recently reported (Hanna et al., 1997). The ovine enzymes have approximately 77% sequence identity with the human enzyme although the two sequences are completely different in the N-terminus comprising 32 residues. This result suggests that pyridoxal kinase is highly homologous in mammalian species.

• KSBB Journal
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• 제16권1호
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• pp.36-40
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• 2001

In order to study the molecular mechanism of $\gamma$-aminobutyric acid (GABA) production in plants, we cloned and sequenced a partial glutamate decarboxylase (GAD) cDNA from the Arabidopsis thaliana cDNA library, using primers targeted at highly conserved sequences of the petunia GAD gene. The cDNA fragment was inserted into TA cloning vector with T7 promoter and the recombinant plasmid obtained was used to transform E. coli. The plasmid DNA purified from the transformed E. coli was digested with EcoRI and the presence of the insert was confirmed. Nucleotide sequence analysis showed that the fragment is a partial Arabidopsis thaliana GAD gene and that the sequence showed 98% and 78% identity to the region of the putative Arabidopsis thaliana GAD sequences deposited in GenBank, Accession nos: U46665 and U10034, respectively. The amino acid sequence deduced from the partial Arabidopsis thaliana GAD gene showed 99% and 91% identities to the GAD sequences deduced from the genes of the U46665 and U10034, respectively. The partial cDNA sequence determined may facilitate the study of the molecular mechanism of GABA metabolism in plants.

• 식물조직배양학회지
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• 제24권2호
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• pp.113-117
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• 1997

Simple sequence repeats (SSR)는 진핵생물체에 널리 산재되어 있으며, 큰 다형현상을 나타내고, polymerase chain reaction (PCR)으로 쉽게 분석된다. 이 연구의 목적은 서로 다른 Chlamydomonas reinhardtii계통간의 다형현상과 Chlamydomonas의 SSR 좌위에서의 유전양상을 결정하는데 있었다. C. reinhardtii의 genomic DNA library를 만들어 $^{32}$P로 라벨링한 (AC)$_{11}$ probe를 이용하여 (CA/GT)n 반복서열을 가지는 clone을 선택하기위해 screen하였다. 선택된 clone을 sequencing하여 (CA/GT)n sequence에 인접한 PCR primer set를 제조하였다. PCR은 여러 C. reinhardtii 계통의 SSR 좌위를 증폭하기 위하여 사용하였다. 그 좌위는 및몇 C. reinhardtii 계통에서 다형현상을 보였다. 그러나 그 좌위에서 C. reinhardtii의 6계통중 4계통만 DNA가 PCR 증폭을 하였고 2계통은 증폭을 하지 않았다. C. reinhardtii와 C. smithii의 교배로 생긴 4배체에서 2:2의 분리비를 보여주었는데, 이는 단순한 멘델의 유전양상을 나타낸다. 이러한 결과로 미루어 SSR 다형현상은 Chlamydomonas의 개체 식별, 개체군 연구, 연쇄 분석, 그리고 유전자 지도 작성을 하는데 유용할 것이다.다.

• Applied Biological Chemistry
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• 제45권3호
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• pp.129-133
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• 2002

키토산 분해물을 시금치와 상추에 살포하였을 때, nitrate 함량이 감소되었으며, 이러한 감소는 nitrate reductase 활성의 증가에 기인한 것으로 나타났다. 이에 따라 채소 중 질산염을 가장 많이 축적하는 채소 중 하나인 시금치의 nitrate reductase를 식물체내에 과량 발현시켜 질산염 축적을 줄이기 위한 연구를 수행하였다. 첫 단계로 시금치 mRNA로부터 RT-PCR을 이용하여 cDNA를 분리, 증폭하고 벡터에 클로닝하여 염기서열을 결정하였다. 시금치 nitrate reductase cDNA의 염기서열은 다른 식물체에서 분리된 nitrate reductase 유전자들과 상당히 높은 상동성($71{\sim}82%$)을 보였고, 이미 발표된 시금치 nitrate reductase cDNA의 염기서열과 비교하였을 때 단지 두 염기만이 달랐다.

• 생명과학회지
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• 제7권3호
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• pp.167-171
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• 1997

번역개시 및 인산화의 조절에 관여하는 RNA와 단백질의 결합 및 인식기작을 연구하기 위해서[$\alpha$^{-32}$P] UMP-labeled HIV Rev-responsive element(RRE) RNA를 이용한 affinity screening에 이해서 Hela ZA-PII cDNA library로부터 이중선RNA결합단백질의 cDNA (RBFII)를 분리하였다. RBFII의 cDNA에 대한 염기서열을 결정하였으며 기존에 연구된 바 있는 RBFII(RBF 또는 TRBP로 보고되었으며 본 연구에서는 RBFII와 구분하기 위해 RBFI으로 명명)과 대부분의 경우 공통적인 ORF를 지니는 것으로 나타났다. 그러나 5’말단에서는 공통적인 ORF 가 RBFI의 경우 21개의 아미노산을 의미하는 63 nt가 Lac-Z의 N-말단에 연결된데 비해서 특이한 46개 아미노기를 의미하는 138nt가 존재함이 밝혀졌다. 5’-말단에 처음 나타나는 ATG 및 부근의 염기서열을 분석해 볼 때 양 cDNA는 5’말단이 완전하지 않은 것으로 사료된다.