• 한국콘텐츠학회:학술대회논문집
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• 한국콘텐츠학회 2007년도 추계 종합학술대회 논문집
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• pp.522-526
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• 2007

중중급성호흡기증후군(SARS, Severe Acute Respiratory Syndrome)은 전 세계적으로 알려진 바가 없었던 신종 급성 전염성 질환으로써, 2003년 아시아로부터 북미와 유럽지역까지 빠른 속도로 전파되어 나간 이후로부터 많은 과학자들의 연구의 대상이 되어오고 있다. 계통발생학적인 관점에서 SARS 바이러스는 Coronavirus 속에 속하는 것으로 알려져 있으나, 전체적인 유전정보 면에서는 다른 코로나바이러스들에 비하여 진화상으로 보존된 부분들이 현저하게 적은 경향을 나타낸다. 자연계에서의 SARS 코로나바이러스(SARS-CoV)의 숙주생물종에 대해서는 아직까지도 명확히 알려지지 않고 있다. 본 연구에서는 SARS-CoV의 유전서열들을 대상으로 다중서열정렬법, 계통발생학적 분석기법 및 다변량 통계분석법 등과 같은 바이오인포매틱스 분석기법들을 활용하여 이 바이러스의 유전정보 패턴을 분석하였다. Relative synonymous codon usage(RSCU)값을 포함하는 여러 유전정보 파라미터들은 Coronavirus와 Lentivirus 속과 Orthomyxoviridae과로부터 수집된 총 30,305개의 암호화 서열들로부터 계산이 되었으며 이 모든 계산은 KISTI 슈퍼컴퓨팅센터의 SMP 클러스터 상에서 수행되었다. 분석 결과, SARS-CoV는 feline 코로나바이러스와 매우 유사한 RSCU 패턴을 나타내었는데, 이것은 기존에 보고되었던 혈청학적인 연구결과와 일치하는 결과였다. 또한 SARS-CoV와 human immunodeficiency virus 및 influenza A virus는 공통적으로 각각이 속한 속이나 과내에서 상대적으로 낮은 RSCU bias를 나타내어서 이와 같은 현상이 이들 바이러스들이 종 간 장벽을 뛰어넘어 전파되는 과정에 영향을 미쳤을 가능성을 시사하였다. 결론적으로 이와 같은 바이오인포매틱스 분석기법들을 활용한 대용량의 유전정보 분석은 유전체 역학 연구에 효과적으로 사용될 수 있을 것으로 기대된다.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.75-87
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• 2021

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp₄₅ signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP_Usp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC₅₀ values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.138-144
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• 1995

The cDNA, encoding human lysozyme (HLY) which was isolated from a human placenta cDNA library, has been well characterized (Yoshimura et al., 1988). Based on the communication, we have prepared an artificial HLY gene from chemically synthesized 38-oligomer with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, an expression vector, pHKl was constructed by inserting the HLY gene, containing a synthetic HLY secretion signal sequence, between the yeast GAP promoter and PH05 terminator. From a lysoplate assay, we have confirmed an yeast transformant harboring a pHK1 which makes a clearing zone on the overlayed Micrococcus luteus. This result means a chemically synthesized HLY gene which was normally expressed and secreted in yeast.

• Journal of Microbiology and Biotechnology
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• 제22권9호
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• pp.1202-1207
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• 2012

A gene, ClCDA, encoding chitin deacetylase from Colletotrichum lindemuthianum, was optimized according to the codon usage bias of Pichia pastoris and synthesized in vitro by overlap extension PCR. It was secretorily expressed in P. pastoris GS115 using the constitutive expression vector pHMB905A. The expression level reached the highest with 110 mg/l culture supernatant after 72 h of methanol induction, which comprised 77.27 U/mg chitin deacetylase activity. SDS-PAGE, mass spectrometry, and deglycosylation assays demonstrated that partial recombinant protein was glycosylated with an apparent molecular mass of 33 kDa. The amino acid sequences of recombinant proteins were confirmed by mass spectrometry.

• Animal Systematics, Evolution and Diversity
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• 제26권3호
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• pp.197-201
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• 2010

We sequenced the whole mitochondrial genome of Dendronephthya gigantea (Anthozoa: Octocorallia: Nephteidae), the first mitochondrial genome sequence report in the Family Nephtheidae. The mitochondrial genome of D. gigantea was 18,842 bp in length, and contained 14 protein coding genes (atp6 and 8, cox1-3, cytb, nd1-6 and 4L, and msh1), two ribosomal RNAs, and only one transfer RNA. The gene content and gene order is identical to other octocorals sequenced to date. The portion of the noncoding regions is slightly larger than the other octocorals (5.08% compared to average 3.98%). We expect that the information of gene content, gene order, codon usage, noncoding region and protein coding gene sequence could be used in the further analysis of anthozoan phylogeny.

• BMB Reports
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• 제41권5호
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• pp.404-407
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• 2008

A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2.

• 한국미생물·생명공학회지
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• 제22권5호
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• pp.477-484
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• 1994

Using a methylotrophic yeast Hansenula polymorpha, a heterologous gene expression and secretion system was developed for the production of hEGF(human Epidermal Growth Factor) which has been shown to promote epithelial cell proliferation and to inhibit gastric acid secretion. The hEGF gene was chemically synthesized according to the preferred codon usage in H. polymor- pha and expressed under the control of the strong and inducible methanol oxidase(MOX) promoter. The mating factor $\alpha$ pre-pro leader sequence of Saccharomyces cerevisiae was employed for hEGF to be secreted into the extracellular medium. This expression cassette was stably integrated into the host chromosomal DNA. Mature hEGF was efficiently expressed and secreted into the extracel- lular medium. About 24 mg/l of hEGF was detected in the cuture supernatant of a transformant with pA-EGF3 under the suboptimal culture conditions.

• Molecules and Cells
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• 제28권3호
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• pp.155-165
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• 2009

The newly sequenced complete mitochondrial genome of the brown marmorated stink bug, Halyomorpha halys($St{\aa}l$) (Hemiptera: Pentatomidae), is a circular molecule of 16,518 bp with a total A+T content of 76.4% and two extensive repeat regions in A+T rich region. Nucleotide composition and codon usage of H. halys are about average when compared with values observed in 19 other published hemipteran mitochondrial genomes. Phylogenetic analyses using these 20 hemipteran mitochondrial genomes support the currently accepted hypothesis that suborders Heteroptera and Auchenorrhyncha form a monophyletic group. The mitochondrial gene arrangements of the 20 genomes are also consistent with our results.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2003년도 제2차 연례학술대회 발표논문집
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• pp.210-219
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• 2003

오페론(operon)은 보통 미생물에서 다수의 인접한 유전자들로 구성된 그룹으로 하나의 유전자처럼 공통된 프로모터에 의해 전사되는 단위이다. 오페론을 구성하는 유전자들은 기능적으로 서로 유사하거나 같은 물질대사경로(metabolic pathway) 상에 존재하는 특징을 지니기 때문에 이들은 중요한 의미를 가지며, 미생물 유전체 분석에서 오페론을 구성하는 유전자들을 예측하는 것은 상당히 중요하다. 오페론을 예측하는 이전 연구들로는 이미 알려진 오페론의 특징인 유전자간 거리나 오페론을 구성하는 평균 유전자 개수 등을 이용하는 방법, 마이크로어레이 발현 실험을 이용한 방법, 전유전체(whole genome)들 간의 보존된 유전자 집합(conserved gene cluster)을 이용한 방법 그리고 물질대사경로를 이용한 방법 등이 있다. 본 논문에서는 COG 기능(function) 거리, 유전자 간의 거리, 코돈 사용빈도(codon usage) 그리고COG 기능 거리와 유전자간 거리를 같이 적용한 방법을 이용하여 오페론 예측을 위한 전처리 모델을 생성하였다 전처리 모델을 E. coli 전유전체에 적용해본 결과, 알려진 오페론들의 약 90%가 이를 포함하였다. 따라서 본 논문에서 제시한 전처리 모델은, 추후 오페론 예측을 위한 좋은 도구로 활용할 수 있을 것이다.

• Journal of Plant Biotechnology
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• 제37권3호
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• pp.243-249
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• 2010

Molecular farming is production of pharmaceutically and industrially important proteins in plants. Plants and plant cell culture systems have been used as bio-factory to produce recombinant proteins such as monoclonal antibodies, enzymes, vaccines, hormones, interleukins, commercial enzymes and etc. The terms molecular farming, biofarming, molecular pharming, phytomanufacturing, recombinant or plant-made industrials, planta-pharma, plant bioreactors, plant biofactory, and pharmaceutical gardening are used interchangeably. Molecular farming can provide safe and inexpensive pharmaceutical proteins as well as commercial ones. In spite of several advantages of molecular farming such as safety and inexpensive cost, there are also a couple of drawbacks in the existing technology. One of them is low expression level of target gene in plants, which has been improved by optimizing gene-based codon usage, screening of strong promoters, expression of transcription factors, subcellular targeting of target proteins, chloroplast transformation, and transient expression using viral expression system (magnifection). Some plant-based commercial proteins have already been in markets and more than twenty plant-based pharmaceuticals have been in clinical trials, from that we can expect that several plant-based pharmaceutical proteins will be seen in the markets in the near future.