• Journal of Microbiology and Biotechnology
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• 제9권1호
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• pp.113-118
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• 1999

The insulin-like growth factor (IGF) is found in all vertebrates and its type-II molecule is regarded as a fundamental embryonic growth factor during development. We have firstly identified, in this study, a cDNA clone corresponding to IGF-II (flIGF-II) from the adult brain of the teleost, Paralichthys olivaceus. We also examined the tissue expression of flIGF-II in several adult tissues by RT-PCR. The flIGF-II cDNA contained a complete ORF consisting of 215 amino acids and one stop codon. Its molecular characteristics appear to be similar to the previously identified IGF-II molecules, in which a common primary structure exhibiting B, C, A, D, and E domains is evidently observed. This cDNA clone seems to be cleaved at $Ala_{52}$ for the $NH_2$-end signal peptide and appears to produce a 98 amino acid-long E-peptide from the $Arg^{118}$. The functional B-D domain regions, therefore, include 65 amino acids and is able to encode a 7.4-kDa protein. The most prominent structural difference between IGF-I and IGF-II was that the D domain of IGF-II exhibits a two-codon-deleted pattern compared to the 8 amino acid-containing IGF-I. The insulin family signature in the A domain and six cysteins forming three disulfide bridges between the B and A domains were evolutionary-conserved from teleosts to mammalian IGF-II. Interestingly, the E-peptide region appears to provide a distinct hallmark between teleosts in amino acid composition. The flIGF-II shows 85.1% of sequence identity to salmon and trout, 90.6% to tilapia, and 98.4% to perch in amino acid level. In tissue expressions of IGF-II, it is very likely that flIGF-II has a significant expression in the adult brain. However, liver seems to be the main source for IGF-II production, and relatively low signals were observed in the adult muscle and kidney. Taken together, it would be concluded that the functional region for IGF-II mRNA is highly similar in phylogeny and is evolutionary, conserved as a mediator for the growth of vertebrates.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2003년도 추계학술발표대회 논문요약집
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• pp.42-42
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• 2003

The far eastern catfish (Silurus asotus) growth hormone (GH) gene was cloned and characterized. The complete nucleotide sequences of genomic GH gene sequences as well as a catfish GH cDNA were obtained by RT-PCR and gene filter screening. The GH cDNA and genomic gene span 1.0 and 1.8 kb from the start codon to the polyadenylation signal, respectively. Both on cDNA and gDNA GH genes, the sequence polymorphism was detected including various silence mutations. The genomic GH gene comprised of only four exons and three introns, which was novel type of fish GH gene structure. The evolutionary relation of the catfish GH gene was inferred based on the comparative phylogenic analysis using the gene structures and sequences.

• Journal of Microbiology
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• 제33권2호
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• pp.165-170
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• 1995

The 356 amino acid long lambda integrase protein of bacteriophage .lambda. constains two autonomous DNA binding domains with distinct sequence specificities. The amino terminal domain of integrase is implicated to bind to the arm-type sequences and the carboxyl domain interacts with the coretype sequencess. As a first step to understand the molecular mechanism of the integrase-DNA interaction at the arm-type site, the int(am)94 gene carrying an amber mutation at the 94th codon of the int was cloned under the control of the P$\_$tac/ promoter and the lacI$\_$q/ gene. The Int(am)94 mutant protein of amino terminal 93 amino acid residues can be produced at high level from a suppressor free strain harboring the plasmid pInt(am)94. The arm-type binding activity of Int(am)94 were measured in vivo and in vitro. A comparison of the arm-type binding properties of the wild-type integrase and the truncated Int(am)94 mutant indicated that the truncated fragment containing 93 amino acid residues carry all the determinants for DNA binding at the arm-type sites.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2747-2751
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• 2014

Background: DNA repair pathways play a crucial role in maintaining the human genome. Previous studies associated DNA repair gene polymorphisms (XPD Lys751Gln, XRCC1 Arg280His and XRCC1 Arg399Gln) with nasopharyngeal carcinoma. These non-synonymous polymorphisms may alter DNA repair capacity and thus increase or decrease susceptibility. The present study aimed to determine the genotype distribution of XPD codon 751, XRCC1 codon 280 and codon 399 polymorphisms and haplotype associations among NPC cases and controls in the Malaysian population. Materials and Methods: We selected 157 NPC cases and 136 controls from two hospitals in Kuala Lumpur, Malaysia for this study. The polymorphisms studied were genotyped by PCR-RFLP assay and allele and genotype frequenci es, haplotype and linkage disequilibrium were determined using SNPstat software. Results: For the XPD Lys751Gln polymorphism, the frequency of the Lys allele was higher in cases than in controls (94.5% versus 85.0%). For the XRCC1 Arg280His polymorphism, the frequency of Arg allele was 90.0% and 89.0% in cases and controls, respectively and for XRCC1 Arg399Gln the frequency of the Arg allele was 72.0% and 72.8% in cases and controls respectively. All three polymorphisms were in linkage disequilibrium. The odds ratio from haplotype analysis for these three polymorphisms and their association with NPC was 1.93 (95%CI: 0.90-4.16) for haplotype CGC vs AGC allele combinations. The global haplotypte association with NPC gave a p-value of 0.054. Conclusions: Our study provides an estimate of allele and genotype frequencies of XRCC1Arg280His, XRCC1 Arg399Gln and XPD Lys751Gln polymorphisms in the Malaysian population and showed no association with nasopharyngeal cancer.

• 미생물학회지
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• 제24권3호
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• pp.236-242
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• 1986

The nucleotide sequence of the genetic control site of Bacillus subtilis gene for $(1-4)-{\beta}-D-glucan$ endoglucanase (cellulase) was determined according to the procedures of the dideoxy chain termination method(Sanger et. al., 1977). The deduced amino acid sequence of this enzyme has a hydrophobic signal peptide at the $NH_2$ terminus similar to those found in fifteen other extracellualr enzymes from Bacillus species. This is followed by a sequence resembling the Bacillus ribosome binding site 14 nucleotide before the first codon of the gene. The presumptive promoter sequence was located 92 base pairs upstream fromthe initiation codon. The homology region in signal sequences was striking when comparing all the signal sequences of sixteen extracellular enzymes from Bacillus species so far compiled.

• BMB Reports
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• 제32권5호
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• pp.436-439
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• 1999

Shigella sonnei is an important cause of human enteric infections. S. sonnei KNIH104S was previously reported to be isolated from Korean shigellosis patients. We cloned a 2.8-kb KpnI fragment containing the recA gene encoding a recombinase from the chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named pRAK28. E. coli HB101, a recA mutant, cannot grow on Luria-Bertani medium in the presence of the alkylating agent methylmethane sulfonate, however, E. coli HB101 harboring pRAK28 was found to grow on this medium. As far as we know, we are the first to sequence the recA gene from S. sonnei. This gene is composed of 1062 base pairs with an ATG initiation codon and a TAA termination codon. Nucleotide sequence comparison of the S. sonnei recA gene exhibited 99.7% and 99.5% identity with those of S. flexneri and E. coli, respectively.

• 미생물과산업
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• 제16권1호
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• pp.7-17
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• 1990

In summary, the nature of the amino acid changes that impart oncogenicity in either the focus formation or tumorigenicity assay can be inferred by analysis of PCR-amplified DNA from tumor-derived NIH3T3 transformants and confirmed by analysis of primary liver tumors. Putative activating mutations in the c-K-ras genetic locus have been shown to involve a single-base modification of either G-C base pair at codon 12 leading to aspartate or cystein substitutions for glycine. The oncogenicity of an N-ras oncogene containing the N-ras C gene region may be related to an amino acid substitution of valine for glycine at codon 13.

• Pediatric Infection and Vaccine
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• 제15권2호
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• pp.195-201
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• 2008

목 적 : 본 연구는 어린 연령의 소아에서 선천성 면역력의 중요한 인자인 Mannose Binding Lectin (MBL2) 유전자 다형성의 분포를 가와사끼병 환자군과 정상대조군에서 분석함으로써, MBL2 유전자의 다양성이 소아의 가와사끼병의 발생에 기여하는지를 밝히고자 하였다. 방 법 : 대상은 2003년 10월부터 2005년 3월까지 18개월 동안 분당서울대학교병원 소아청소년과에서 가와사끼병으로 진단받고 치료받은 112명의 환자군과 건강한 성인 224명의 대조군으로 선정하였다. 유전형 분석은 선천성 면역의 중추적 역할을 하는 MBL2 유전자의 codon 54의 분포를 환자대조군에서 비교하여 분석하였다. 결 과 : 환자군 112명의 중앙 연령은 27개월로, 24개월 미만이 45.5%, 24개월 이상이 54.5%를 차지하였다. MBL2 codon 54 유전형은 환자군과 대조군에서 모두 Hardy-Weinberg 평형을 이루었다. 환자군에서 MBL2 유전형은 유전형 aa 67.9%, 이형접합 ab 29.5%, 그리고 유전형 bb 2.6%로 나타났다. Codon 54의 다형성은 환자군과 대조군간에 차이가 없었다. 또한 연령 24개월을 기준으로 하여 비교한 유전형의 분포에도 차이가 없는 것으로 나타났다. 결 론 : 본 연구 결과, 한국인 소아에서 MBL2 유전자의 다형성이 가와사끼병의 발병에 기여하는 유전적 인자임을 밝히지 못하였다. 향후 전향적으로 수집한 더 많은 수의 환자군과 적절한 대조군을 통한 연구가 시행 되어야 할 것으로 생각한다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.736-738
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• 2003

Ornithine decarboxylase (ODC) is the key enzyme in the synthetic pathway of polyamines. This enzyme is a homodimeric and a pyridoxal 5-phosphate (PLP) dependent enzyme. We have isolated, a cDNA clone encoding ODC from brain cDNA library constructed from flounder (Paralichthys olivaceus). The ODC cDNA contained a complete ORF consisting of 460 amino acids and one stop codon with comparison to nucleotide sequences of the flounder, zebrafish and rat ODC genes, the ODC genes were highly conserved. The transcription of ODC was analyzed with reverse transcription-polymerase chain reaction (RT-PCR) species in brain, kidney, liver, and embryo.

• 한국미생물·생명공학회지
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• 제43권1호
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• pp.1-8
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• 2015

돼지생식기호흡기증후군(porcine reproductive and respiratory syndrome; PRRS) 바이러스의 ORF5a 단백질이 바이러스 생장에 필수적인 단백질인지 확인하기 위해서 PRRS 바이러스 감염성 클론을 이용하여 ORF5a 단백질 유전자를 결손시킨 변이 클론을 제작하였다. 야생형 PRRS 바이러스 감염성 클론과 ORF5a 단백질이 결손된 변이 클론을 BHK-tailless pCD163 세포에 transfection시킨 결과 변이클론에서감염성있는바이러스가숙주세포로부터만들어지지않았다. 이결과가 ORF5a 단백질발현의부재때문인지검증하기위해서 BHK-tailless pCD163-tailless 세포에 ORF5a 단백질을안정적으로발현하는세포주를제작하였고이세포주에동일한 transfection 실험을한결과세포에서공급되는 ORF5a 단백질발현에의해감염성있는바이러스가만들어지는것을확인하였다. 이로써 ORF5a 단백질이 PRRS 바이러스가 생장하는데 있어서 필수적인 단백질임을 확인할 수 있었다.