• 제목/요약/키워드: cDNA sequence

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cDNA Sequence and mRNA Expression of a Novel Peroxiredoxin from the Firefly, pyrocoelia rufa

  • Jin, Byung-Rae;Lee, Kwang-Sik;Kim, Seong-Ryul;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제4권2호
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    • pp.101-107
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    • 2002
  • We describe here the cDNA sequence and mRNA expression of a novel family of the antioxidant protein, peroxiredoxin, from the firefly, Pyracoetia ruin. The 555 bp cDNA sequence codes for a 185 amino acid protein with a calculated molecular mass of approximately 21 kDa. The deduced protein of P. rufa peroxiredoxin gene contains two conserved cysteine residues. Alignment of the deduced protein of P. rufa peroxiredoxin gene showed 71.1% protein sequenceidentity to known insect Drosophila melanogaster peroxiredoxin. Northern blot analysis revealed that the P. rufa peroxiredoxin is specifically expressed in the fat body of P. rufa larvae.

cDNA Sequence and mRNA Expression of a Novel Serine Protease from the Firefly, Pyrocoelia rufa

  • Lee, Kwang-Sik;Kim, Seong-Ryul;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제5권1호
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    • pp.103-108
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    • 2002
  • We describe here the cDNA sequence and mRNA expression of a novel serine pretense from the firefly, Pyrocoelia rufa. The 771 bp cDNA encodes for 257 amino acid residues. The deduced protein of P. rufa serine pretense gene contains the catalytic triad and six-conserved cysteine residues. Alignment of the deduced protein of P. rufa serine pretense gene showed 47.4% protein sequence identity to known coleopteran insect Rhyzopertha dominica midgut trpsin-like enzyme. Northern blot analysis revealed that the P. rufa serine pretense is specifically expressed in the midgut of P. rufa larvae.

Molecular Analysis of Complete SSU to LSU rDNA Sequence in the Harmful Dinoflagellate Alexandrium tamarense (Korean Isolate, HY970328M)

  • Ki, Jang-Seu;Han, Myung-Soo
    • Ocean Science Journal
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    • 제40권3호
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    • pp.155-166
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    • 2005
  • New PCR primers (N=18) were designed for the isolation of complete SSU to LSU rDNA sequences from the dinoflagellate Alexandrium tamarense. Standard PCR, employing each primer set selected for amplifications of less than 1.5 kb, successfully amplified the expected rDNA regions of A. tamarense (Korean isolate, HY970328M). Complete SSU, LSU rDNAs and ITS sequences, including 5.8S rDNA, were recorded at 1,800 bp, 520 bp and 3,393 bp, respectively. The LSU rDNA sequence was the first report in Alexandrium genus. No intron was found in the LSU rRNA coding region. Twelve D-domains within the LSU rDNA were put together into 1,879 bp (44.4% G+C), and cores into 1514 bp (42.8% G+C). The core sequence was significantly different (0.0867 of genetic distance, 91% sequence similarity) in comparison with Prorocentrum micans (GenBank access. no. X16108). The D2 region was the longest in length (300 bp) and highly variable among the 12 D-domains. In a phylogenetic analysis using complete LSU rDNA sequences of a variety of phytoplankton, A. tamarense was clearly separated with high resolution against other species. The result suggests that the sequence may resolve the taxonomic ambiguities of Alexandrium genus, particularly of the tamarensis complex.

Cloning and Expression of Alkaline Phosphatase Gene from Schizosaccharomyces pombe

  • Kang, Sung-Won;Cho, Young-Wook;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • 제34권3호
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    • pp.262-267
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    • 2001
  • A cDNA coding alkaline phosphatase (AP) homologue was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The nucleotide sequence of the cloned cDNA appeared to lack the N-terminal coding region. The genomic DNA encoding alkaline phosphatase homologue was isolated from S. pombe chromosomal DNA using PCR. The amplified DNA fragment was ligated into plasmid pRS315 to generate the recombinant plasmid pSW20. The DNA insert was subcloned as two smaller fragments for nucleotide sequencing. The sequence contains 2,789 by and encodes a protein of 532 amino acids with a molecular mass of 58,666 daltons. The S. pombe cells containing plasmid pSW20 showed much higher AP activity compared with the yeast cells with vector only This indicates that the cloned AP gene apparently encodes AP The predicted amino acid sequence of the S. pombe AP shares homology with those of other known APs.

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Analysis of Partial cDNA Sequence from Human Fetal Liver

  • Kim, Jae-Wha;Song, Jae-Chan;Lee, In-Ae;Lee, Young-Hee;Nam, Myoung-Soo;Hahn, Yoon-Soo;Chung, Jae-Hoon;Choe, In-Seong
    • BMB Reports
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    • 제28권5호
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    • pp.402-407
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    • 1995
  • Single-run Partial cDNA sequencing was conducted on 1,592 randomly selected human fetal liver cDNA clones of Korean origin to isolate novel genes related to liver functions. Each partial cDNA sequence determined was analyzed by comparing it with the databases. GenBank, Protein Information Resource (PIR) and SWISS-PROT Protein Sequence Data Bank. From a set of 1.592 cDNA clones reported here, 1,433 (90.0% of the total) were informative cDNA sequences. The other 159 clones were identified as DNA sequences which had originated from the cloning vector. Among 1,433 informative partial cDNA sequences, 851 (59.3%) clones were revealed to be identical to known human genes. These known genes have been classified into 225 different kinds of genes. In addition, 340 clones (23.7%) showed various degrees of homology to previously known human genes. Ninety four (6.6%) clones contained various repeated sequences. Twenty four (1.7%) partial cDNA sequences were found to have considerable homology to known genes from evolutionarily distant organism such as yeast, rice, Arabidopsis, mouse and rat, based on database matches, whereas 124 (8.7%) had no Significant matches. Human homologues to functionally characterized genes from different organisms could be classified as candidates for novel human genes of similar functions. Information from the partial cDNA sequences in this study may facilitate the analysis of genes expressed in human fetal liver.

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연속적 차분 확장 기반 가역 DNA 워터마킹 (Consecutive Difference Expansion Based Reversible DNA Watermarking)

  • 이석환;권기룡
    • 전자공학회논문지
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    • 제52권7호
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    • pp.51-62
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    • 2015
  • 대용량의 DNA 정보 저장, DNA 서열 저작권 보호를 위한 DNA 워터마킹, 및 비밀 통신을 위한 DNA 스테가노그라픽에 대한 관심이 증대되면서, 원본 DNA 서열의 기능 유지와 복원이 가능한 가역성 DNA 워터마킹이 필요하다. 본 논문에서는 비부호영역 DNA 서열을 이용한 DE(Difference expansion) 기반 가역 DNA 워터마킹 기법을 제안한다. 가역 DNA 워터마킹에서는 생물학적 기능 변경이 없고, 문자 형태의 서열 내에 대용량의 데이터를 은닉하여야 하며, 원본 DNA 서열이 복원되어야 한다. 제안한 방법에서는 문자 서열을 십진수 형태의 수치계수로 변환한 다음, 인접 수치 계수 쌍의 DE 기반 다중비트 은닉 방법(DE-MBE, DE based multiple bits embedding)과 이전 은닉 수치계수를 예측으로 한 연속 DE 기반 다중비트 은닉 방법들(C-DE-MBE, consecutive DE based multiple bits embedding)에 의하여 워터마크가 은닉된다. 은닉 과정에서는 워터마크된 서열에 의하여 부호영역을 나타내는 허위 시작코돈 발생을 방지하기 위하여 비교 탐색을 수행한다. 실험 결과로부터 제안한 방법이 기존 방법에 비하여 높은 은닉 용량을 가지며, 허위 시작코돈이 발생되지 않으며, 기준 서열없이 원본 DNA 서열이 복원됨을 확인하였다.

DNA Structural Perturbation Induced by the CPI-Derived DNA Interstrand Cross-linker : Molecular Mechanisms for the Sequence Specific Recognition

  • Park, Hyun-Ju
    • Archives of Pharmacal Research
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    • 제24권5호
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    • pp.455-465
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    • 2001
  • The highly potent cytotoxic DNA-DNA cross-linker consists of two cyclopropa[c]pyrrolo[3,4-3]indol-4(5H)-ones insoles [(+)-CPI-I] joined by a bisamido pyrrole (abbreviated to "Pyrrole"). The Pyrrole is a synthetic analog of Bizelesin, which is currently in phase II clinical trials due to its excellent in vivo antitumor activity. The Pyrrole has 10 times more potent cytotoxicity than Bizelesin and mostly form DNA-DNA interstrand cross-links through the N3 of adenines spaced 7 bp apart. The Pyrrole requires a centrally positioned GC base pair for high cross-linking reactivity (i.e., $5^1$-T$AT_2$A*-$3^1$), while Bizelesin prefers purely AT-rich sequences (i.e., $5^1$-T$AT_4$A*-$3^1$, where /(equation omitted) represents the cross-strand adenine alkylation and A* represents an adenine alkylation) (Park et al., 1996). In this study, the high-field $^1$H-NMR and rMD studies are conducted on the 1 1-mer DNA duplex adduct of the Pyrrole where the 5′(equation omitted)TAGTTA*-3′sequence is cross-linked by the drug. A severe structural perturbation is observed in the intervening sequences of cross-linking site, while a normal B-DNA structure is maintained in the region next to the drug-modified adenines. Based upon these observations, we propose that the interplay between the bisamido pyrrole unit of the drug and central C/C base pair (hydrogen-bonding interactions) is involved in the process of cross-linking reaction, and sequence specificity is the outcome of those interactions. This study suggests a mechanism for the sequence specific cross-linking reaction of the Pyrrole, and provides a further insight to develop new DNA sequence selective and distortive cross-linking agents.

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Arabidopsis thaliana로부터 지방산 불포화효소 유전자의 분석 (Characterization of a fad3 cDNA Encoding Microsomal Fatty Acid Desaturase from Arabidopsis thaliana)

  • 박희성;임경준
    • 식물조직배양학회지
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    • 제24권2호
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    • pp.93-97
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    • 1997
  • Linoleic acid를 linolenic acid로 전환시키는 지방산불포화효소의 유전자(fad3)를 유채의 fad3 DNA probe를 이용한 plaque hybridization방법으로 $\lambda$ZAPII Arabidopsis thaliana cDNA expression library로부터 분리하였으며 1.8 kb-EcoRI DNA조각을 지니는 lambda clone을 pGEM7으로 subcloning하여 염기서열을 분석하였다. 그 결과로부터의 아미노산서열분석에 의하면 fad3 유전자는 open reading frame이 386개의 아미노산으로 이루어졌으며 44,075 Da의 분자량이 예측되고 있다. 엽록체의 $\omega$-3 지방산불포화효소(fad7)와 endoplasmic reticulum의 지방산불포화효소(fad2)와의 비교시 각각 70%와 58%의 유사성이 나타났다. 특히 82-151의 아미노산서열과 276-333의 아미노산지역은 보존성이 높았으며 이는 불포화지방산효소의 기능에 필수적인 지역으로 보여진다. 한편 대장균을 이용한 IPTG에 의한 Fad3단백질의 유도생성은 대장균의 생육에 독성효과를 나타내는 것으로 나타났다.

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더덕의 주근에서 유래한 Dehydrin 1 (Dhn1) 유전자의 분리 및 분석 (Isolation and Characterization of Dehydrin 1 (Dhn1) gene from Codonopsis lanceolata)

  • 이강;양덕춘
    • 한국자원식물학회지
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    • 제16권3호
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    • pp.238-244
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    • 2003
  • 더덕 뿌리에서 유래한 EST 라이브러리로부터 dehydrin 유전자와 높은 상동성 을 나타내는 full clone cDNA를 얻었다. 더덕의 dehydrin, ClDhn1은 893 bp의 cDNA로 159개의 아미노산을 코딩하는 480 bp의 ORF를 가지고 있다. ClDhn1의 아미노산을 분석해 보면, 전체적으로 높은 친수성을 나타내며, lysine이 풍부한 K 반복구간(KIKEKLPG)을 카르보닐기 쪽에 2개 가지고 있다. 또한, 여러 dehydrin들의 공통적인 특징인 7개의 연속적인 serine잔기가 첫 번째 K반복 구간 앞에 위치한다. 그러나, 아미노기쪽의 DEYGNP보존 구간은 변형(DEHGNP)되어 있다. ClDhn1 유전자는 전사 단계에서 더덕의 뿌리에서 가장 높은 발현 양상을 보이며, 줄기와 잎에서는 적은 양이 발현되었다.