• International Journal of Industrial Entomology and Biomaterials
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• 제3권2호
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• pp.163-168
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• 2001

We isolated and sequenced a cDNA clone corresponding to apolipophorin-III (apoLp-III) from the fall webworm, Hyphantria cunea. The cDNA for apoLp-III codes fer a 187-residue protein (561 bp) with a predicted molecular mass of 20 kDa. The calculated isoelectric point is 8.76. Multiple alignment analysis of the amino acid sequence revealed that H. cunea apoLp-III is most similar to that of Spodoptera litura (71.5% identity), followed by that of Manduca sexta (69.7% identity). They share five amphipathic $\alpha$-helices that are proposed to play a critical role in the binding of apoLp-III to lipophorin.

• 대한의생명과학회지
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• 제2권2호
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• pp.231-240
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• 1996

인체 혈액 응고 9인자는 간에서 생성되며 461개의 아미노산으로 구성된 당 단백질이다. 따라서 인체 혈액 응고 9인자 cDNA를 찾기 위해 태아의 간(fetal liver) cDNA library를 PCR(Polymerase Chain reaction) 방법으로 screening하였으며, 그 결과 ATG개시 코돈으로부터 TAA종료 코돈까지 포함하는 1.4 kb의 9인자 cDNA를 찾았다. 또한 클론된 9인자 cDNA를 박테리아에서 발현시키기 위해 박테리아 발현 벡터인 pGEX-2T 플라스미드에 클로닝하므로써 pGEX-F9 플라스미드를 제조하였다. pGEX-F9로 형질전환된 E. coli에서 PGEX-F9의 발현을 유도하면 73 kDa 크기의 GST-factor9 융합 단백질이 다량생성되며 , 이 단백질이 혈액 응고 9인자 단백질을 함유하는 융합 단잭질임을 혈액 응고 9인자 항체를 이용한 Western blot으로 입증하였다. E. coli에서 발현된 GST-factor 9 융합 단백질은 전체 단백질의 약 20%를 차지하며 GST agarose bead를 이용한 one step purificarion 방법을 통해 GST-factor9 융합 단백질을 쉽게 분리 할 수 있다.

• Genomics & Informatics
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• 제10권3호
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• pp.206-211
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• 2012

DNA barcoding has been widely used in species identification and biodiversity research. A short fragment of the mitochondrial cytochrome c oxidase subunit I (COI) sequence serves as a DNA bio-barcode. We collected DNA barcodes, based on COI sequences from 156 species (529 sequences) of fish, insects, and shellfish. We present results on phylogenetic relationships to assess biodiversity the in the Korean peninsula. Average GC% contents of the 68 fish species (46.9%), the 59 shellfish species (38.0%), and the 29 insect species (33.2%) are reported. Using the Kimura 2 parameter in all possible pairwise comparisons, the average interspecific distances were compared with the average intraspecific distances in fish (3.22 vs. 0.41), insects (2.06 vs. 0.25), and shellfish (3.58 vs. 0.14). Our results confirm that distance-based DNA barcoding provides sufficient information to identify and delineate fish, insect, and shellfish species by means of all possible pairwise comparisons. These results also confirm that the development of an effective molecular barcode identification system is possible. All DNA barcode sequences collected from our study will be useful for the interpretation of species-level identification and community-level patterns in fish, insects, and shellfish in Korea, although at the species level, the rate of correct identification in a diversified environment might be low.

• 한국해양바이오학회지
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• 제1권2호
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• pp.119-125
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• 2006

어류의 insulin-like growth factor-I (IGF-I)의 생화학적 작용기작을 연구하기 위하여 한국산 조피볼락의 IGF-I cDNA 유전자 cloning을 행하였다. 완전한 cDNA 유전자 염기서열은 PCR과 RACE 방법을 통하여 얻어진 DNA로부터 결과를 얻을수 있었다. 결정된 IGF-I의 염기서열은 flounder, chinook salmon, human IGF-I의 염기서열과 비교한 결과 각각 93.6%, 90.7%, 85.4%의 높은 상동성을 보였다. 생화학적으로 활성이 있는 재조합 IGF-I을 얻기 위하여 IGF-I의 B-C-A-D domain 부분을 PCR로 얻은 뒤 E. coli BL21(DE3)에 넣어 overexpression 시켰다. Ni-NTA colummn을 사용하여 순수한 재조합 단백질을 정제할수 있었다. 정제된 단백질은 SDS-PAGE 상에서 7 kDa의 단일 band를 보여 주었으며 [$^3H$]-thymidine 결합정도를 측정하는 방법으로 활성을 가지고 있음을 확인할수 있었다.

• 한국미생물·생명공학회지
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• 제28권1호
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• pp.21-25
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• 2000

We have got several clones from Serratia marcescens which stimulated the cells to use maltose as a carbon source in Escherichia. coli TP2139 ( lac, crp). One of the cloned genes, pCKB17, was further analyzed. In order to find whether the increased expression of the gent was under the direction of maltose metabolism, we constructed several recombinant subclones. We have found that the clone, pCKB17AV, codes maltose metabolism stimulation(mms) gene. E. coli transformed with the cloned gene showed increase in the activity of maltose utilzation, The recombinant proteins expressed by multicopy and induction with IPTG, one polypeptide of 29-kDa, was confirmed by SDS-PAGE. The overexpression of maltose-binding proter protein in the presence of mms gene was confirmed by Western blot analysis. Southern hybridization analysis confirmed that the cloned DNA fragment was originated from S. marcescens chromosomal DNA.

• Journal of Plant Biology
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• 제38권2호
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• pp.137-141
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• 1995

We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

• Journal of Plant Biology
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• 제32권2호
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• pp.67-78
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• 1989

Hybridization of DNA isolated from leaves of Russet Burbank potato with tomato cDNA as a probe revealed the presence of about ten inhibitor 1 genes in the genome. Screening of a genomic library of Russet Burbank potato resulted in isolation of seven different genomic clones carrying inhibitor I genes. One of the genomic clones, clone 2, contained two EcoRI fragments of 3.4 and 1.8 kb in size, respectively, which were hybridized with the probe. The nucleotide sequence of parts of the hybridizing EcoRI fragments revealed that they contain a complete gene which codes for an open reading frame of 107 amino acids. It is interrupted by two intervening sequences of 502 and 493 bp, situated at the positions of codons 17 and 43, respectively, of the open reading frame. Putative regulatory sequences, TATAAA and CCACT, were found at the 5' flanking region. In addition, a copy of a 100 bp repeat found at a tomato inhibitor I gene was identified.

• 한국멀티미디어학회논문지
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• 제16권3호
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• pp.318-329
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• 2013

본 논문에서는 DNA 시퀀스의 불법 복제 및 변이 방지를 위한 DNA 워터마킹 기법을 제안한다. 제안한 DNA 워터마킹은 랜덤 맵핑 테이블에 의하여 코돈들을 랜덤 원형 각도로 수치화한 다음, 웨이블릿 국부계수 최대치의 Lipscihtz regularity 상수에 의하여 삽입 대상 코돈들을 탐색한다. 워터마크 삽입과정에서 DNA의 아미노산 코드가 변경되지 않도록 하기위하여 삼중 코돈들의 랜덤 코돈 원형 각도에 워크마크를 삽입한다. 삽입 대상 코돈들의 길이와 위치는 랜덤 맵핑 테이블에 의존하므로, 이 테이블을 알지 못할 경우, 워터마크 추출이 어렵다. 그리고 제안한 방법은 다양한 길이의 DNA 서열에 64개 코돈(종료, 개시 코돈포함)들의 랜덤 맵핑 테이블을 적용함으로써 동일한 길이의 워터마크 키를 적용한다. 본 실험에서는 랜덤 맵핑 테이블과 삽입 위치의 높은 엔트로피를 통하여 워터마크의 보안성을 확인하였다. 또한 기존의 DNA-Crypt 워터마킹과의 유사한 용량 하에서 제안한 방법이 낮은 염기 변화율을 가지며, 포인트 변이, 삽입 및 삭제 변이에 대하여 낮은 에러률를 가지며, ROC 분석을 통하여 우수한 검출 능력을 가짐을 확인하였다.

• BMB Reports
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• 제39권5호
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• pp.578-585
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• 2006

The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMP-binding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.

• Fisheries and Aquatic Sciences
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• 제9권1호
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• pp.22-29
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• 2006

We determined the partial cDNA of Type II procollagen ${\alpha}1$[pro-${\alpha}1$(II)] chain (1802 bp) of the skate Raja kenojei, which codes 581 amino acid residues. The partial structure of the pro-${\alpha}1$(II) chain consisted of a part of triple helical region (309 residues) and a C-domain (272 residues). Comparing the chain to other vertebrates showed relatively low homology (about 50%) at the amino acid level. However, eight Cys residues in the C-domain of the skate pro-${\alpha}1$(II) chain were conserved in common with those of other vertebrates. The skate pro-${\alpha}1$ (II) chain mRNA was detected by RT-PCR of various tissues, but was undetected in tissues containing Type II collagen. The low homology and unexpected expression pattern suggest the presence of another mRNA variant of the skate pro-${\alpha}1$(II) chain. The present study is the first report of the primary structure of pro-${\alpha}1$(II) chain in an elasmobranch.