• BMB Reports
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• 제39권4호
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• pp.346-354
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• 2006

The full-length cDNA of grass carp (Ctenopharyngodon idellus) and silver carp (Hypophthalmichthys molitrix) uncoupling protein 2 (UCP2) was obtained from liver. The grass carp UCP2 cDNA was determined to be 1152 bp in length with an open reading frame that encodes 310 amino acids. Five introns (Intron 3, 4, 5, 6 and 7) in the translated region, and partial sequence of Intron 2 in the untranslated region of grass carp UCP2 gene were also obtained. Gene structure comparison between grass carp and mammalian (human and mouse) UCP2 gene shows that, the UCP2 gene structure of grass carp is much similar to that of human and mouse. Partial UCP2 cDNA sequences of bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella), were further determined. Together with the common carp (Cyprinus carpio) UCP2 sequence from GenBank (AJ243486), multiple alignment result shows that the nucleotide and amino acid sequences of the UCP2 gene, were highly conserved among the five major Chinese carps that belong to four subfamilies. Using beta-actin as control, the ratio UCP2/beta-actin mRNA (%) was determined to be $149.4{\pm}15.6$ (common carp), $127.4{\pm}22.1$ (mud carp), $96.7{\pm}12.7$ (silver carp), $94.1{\pm}26.8$ (bighead carp) and $63.7{\pm}16.2$ (grass carp). The relative liver UCP2 expression of the five major Chinese carps, shows a close relationship with their food habit: benthos and detrituseating fish (common carp and mud carp) > planktivorious fish (silver carp and bighead carp) > herbivorious fish (grass carp). We suggest that liver UCP2 might be important for Chinese carps to detoxify cyanotoxins and bacteria in debris and plankton food.

• Mycobiology
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• 제40권2호
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• pp.107-110
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• 2012

Genes encoding the cellobiohydrolase enzyme (CBHI), designated as cbhI, were isolated from the basidiomycetes Auricularia fuscosuccinea, Pleurotus giganteus, P. eryngii, P. ostreatus, and P. sajor-caju. Initially, the fungal genomic DNA was extracted using a modified cetyltrimethyl ammonium bromide (CTAB) protocol and used as a DNA template. The cbhI genes were then amplified and cloned using the pGEM-T Easy Vector Systems. The sizes of these PCR amplicons were between 700~800 bp. The DNA sequences obtained were similar showing high identity to the cbhI gene family. These cbhI genes were partial consisting of three coding regions and two introns. The deduced amino acid sequences exhibited significant similarity to those of fungal CBHI enzymes belonging to glycosyl hydrolase family 7.

• The Plant Pathology Journal
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• 제18권1호
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• pp.6-11
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• 2002

An isolate of Cucumber mosaic virus (CMV), PaFMl-CMV causing malformation on the fruit of paprika (Capsicum annuum var, grossum) was characterized based on biological reactions, serological relationships, and partial nucleotide sequence analyses. PaFMl-CMV was distinguishable from other isolates of CMYI Mf-(subgroup I) and LS-CMV (subgroup II), in terms of its reactions to some host plants. Polyclonal antibody against PaFMl-CMV showed homologous antigenic relationship with LS-CMV, however, the antibody formed a spur between PaFMl- and Mf-CMV, In the comparison of molecular size of dsRNAs of PaFMl-CMV with Mf- and LS-CMV, PaFMl-CMV had a slightly smaller RNAl and larger RNA2, RNA3, and RNA4. When the CDNA product of PaFMl-CMV coat protein (CP) gene was digested with some restriction enzymes, the fragment pattern was identical with that of LS-CMV The nucleotide and amino acid sequences of PaFMl-CMV CP gene were 99.5% and 98.6% identical with LS-CMV respectively. The data indicate that PaFMl-CMV belongs to subgroup II of CMV, which is the first report in Korea.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.121-121
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• 1998

Artemisia annua, an indigenous plant in Korea, contains a clinically important potent antimalarial principle, artemisinin. Artemisinin is a cadinane-type sesquiterpene endoperoxide. Cadinane synthase catalyzes the first committed step in artemisinin biosynthesis by cyclizing farnesyldiphosphate. In hopes of finding a cadinane synthase gene involved in artemisinin biosynthesis, oligonucleotides were synthesized on. the basis of the consensus nucleotide sequences and Nco I restriction sites for convenience in cloning. Specifically, nucleotide sequences of two highly conserved regions were deduced from the genes of similar function of Hyoscyamus muticus, Nicotiana tabacum, Abies grandis, Lycopersicon esculentum, and Gossypium hirsutum to construct a set of primers for polymerase chain reation (PCR). A 184 bp fragment was found to be amplified by PCR, and subsequently cloned. The gene revealed 62.8% identity in nucleotide and 55.6% in amino acid sequence to correspondent gene of N. tabacum. The gene was different from another sesquiterpene cyclase gene of A. annua, germacranadiene synthase gene, recently reported by Mercke and Bordelius (1998).

• Animal Systematics, Evolution and Diversity
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• 제16권2호
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• pp.203-211
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• 2000

한국에서 양식되어지고 있는 한국산 굴류 4종, 굴(Crassostrea gigas Thunberg), 바위굴(C. nippona Seki), 강굴(C. ariakensis Fujita et Wakiya), 토굴(Ostrea denselamellosa Lischke)의 유전적 근연관계를 조사하고자 미토콘드리아 DNA의 16S rDNA와 cytochrome c oxidase I (COI) 유전자 일부분의 염기서열을 분석하였다. 16S rDNA의 319 bp와 COI유전자의 710 bp를 PCR 증폭하여 염기서열을 결정하였으며, 염기서열과 아미노산서열을 자료로 하여 UPGMA와 neighbor-joining 방법으로 계통수를 작성하고, 종간 유연관계를 확인하였다. Crassostrea 속과 Ostrea 속간 비교에서는 뚜렷한 유전적 분화를 나타내었으며 계통분석 결과, neighbor-joining 방법에 의한 COI의 아미노산 서열분석에서는 굴과 강굴이 자매군을 형성하는 양상을 보였으나 두 유전자의 염기서열과 A+T 비율 비교에서는 굴과 바위굴이 자매군을 형성하는 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제30권4호
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• pp.312-319
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• 2002

저온성 균인 Sporosarcina psychrophilia의 염색체 DNA를 추출하여 Sau3AI으로 부분 절단하고 pUC19 vector에 ligation 시킨 후, Escherichia coli pyrB mutant 균주에 형질전환하여 uracil이 없는 AB배지에서 생존하는 균주를 선택한 후, 그 plasmid를 분리하여 pSMI과 pSM2라고 명명하였다. 두 plasmid의 염기배열을 결정한 결과 pSM2 insert DNA는 pSMl insert DNA 부분을 포함하는 2,606 nucleotide 단편이었다. 염기서열을 분석하였을 때 이것은 1개의 완전한 open reading frame(ORF)과 2개의 부분 ORFs를 포함하고 있었다. 두 번째 위치한 완전한 ORF는 Bacillus caldolyticus aspartate transcarbamylase(pyrB)와 아미노산 서열 수준에서 59% 상동성을 보였고, 첫 번째와 세 번째 위치한 부분적 ORFs는 각각 Bacillus속의 uracil permease(pyrP)와 dihydoorotase(pyrC)와 높은 상동성을 보였다. 그리고 pyrB와 pyrP사이에 intergenic 부분에는 잠재적인 terminator, antiterminator, anti-antiterminator 구조를 포함하고 있었다. 이러한 결과는 S. psychrophilia pyrimidine 생합성에 관련된 유전자들은 다른 Bacillus속에서 알려진 바와 같이 유전자군을 형성하고 있을 것으로 추정했다. S. psychrophilia pyrB 유전자의 생성물을 과다발현 시키고 정제해서 그 단백질을 SDS-PAGE로 확인한 결과 27 kDa 부근에서 band를 확인할 수 있었으며, 정제한 단백질도 ATCase 효소활성을 지니고 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권8호
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• pp.1169-1173
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• 2007

The protein encoded by SLC27A2 gene is an isozyme of long-chain fatty-acid-coenzyme A ligase family, and it converts free long-chain fatty acids into fatty acyl-CoA esters, and thereby plays a key role in lipid biosynthesis and fatty acid degradation. In the present study, SLC27A2 located on human chromosome 15 was selected as candidate gene and we isolated and cloned partial fragments of mRNA sequence and genomic fragments of porcine SLC27A2 gene. The coding region of the gene as determined by alignments shared 90% and 82% identity with human and mouse cDNAs, respectively. Detection in LargeWhite and Meishan breeds showed that a single nucleotide polymorphism (SNP) ($A{\rightarrow}G$) existed in exon 7, which caused corresponding amino acid changed for encoding. In LargeWhite pigs it encoded for Val while in Meishan pigs it encoded for Ile, so we developed the PCR-RFLP genotype method for detection of this polymorphism. Association study in 135 $F_2$ reference family indicated that significant correlation existed between the polymorphism and growth and carcass traits.

• 한국미생물·생명공학회지
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• 제45권2호
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• pp.155-161
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• 2017

Paenibacillus woosongensis의 유전체 부분 염기서열로부터 유추된 xylanase 유전자를 PCR 증폭하여 클로닝하고 염기서열을 결정하였다. 클로닝된 xylanase 유전자는 xyn11B로 명명되었으며, 356 아미노산으로 구성된 단백질을 코드하는 1,071 뉴클레오티드로 이루어졌다. Xyn11B의 아미노산 배열을 분석한 결과 glycosyl hydrolase family 11에 속하는 xylanase와 상동성이 높은 활성영역과 탄수화물 결합영역을 포함하고 있는 다영역 효소로 확인되었다. SignalP4.1 server로부터 아미노 말단의 26개 잔기가 signal peptide로 예측되었다. DEAE-Sepharose와 Phenyl-Separose 컬럼 크로마토그래피 과정을 통해 xyn11B 유전자를 함유한 재조합 대장균의 균체 파쇄상등액으로부터 Xyn11B를 부분 정제하였다. 부분 정제된 Xyn11B의 반응특성을 조사한 결과 pH 6.5와 $50^{\circ}C$에서 최대 반응활성을 보였고 birchwood xylan이나 oat spelt xylan보다 arabinoxylan에 대한 활성이 높았으며 셀룰로스, 만난과 para-nitrophenyl-${\beta}$-xylopyranoside에 대해서는 분해활성이 없었다. Xyn11B의 활성은 $Ca^{2+}$과 $Mg^{2+}$에 의해서는 약간 증가한 반면에 $Cu^{2+}$, $Ni^{2+}$, $Fe^{3+}$, $Mn^{2+}$에 의해서는 크게 저해되었고 SDS에 의해서 완전히 저해되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권3호
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• pp.324-328
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• 2006

Carbonic anhydrase III (CA3) is a member of a multigene family that encode carbonic anhydrase isozymes. In this study, a complete coding sequence of the pig CA3 gene which encodes a 260 amino-acid protein was determined. The amino acid comparison showed high sequence similarities with previously identified human (86.5%) CA3 gene and mouse (91.5%) Car3 gene. The partial genomic DNA sequences were also investigated. The length of intron 1 was 727 bp. Comparative sequencing of three pig breeds revealed that there was a T${\rightarrow}$C substitution at position 363 within intron 1. The substitution was situated within a NcoI recognition site and was developed as a PCR-restriction fragment length polymorphism (RFLP) marker for further use in population variation investigations and association analysis. Two alleles (A and B) were identified, and 617 bp fragments were observed for the AA genotype and 236 bp and 381 bp fragments for the BB genotype. The polymorphism of CA3 was detected in 8 pig breeds. Allele B was predominant in the Western pig breeds. In addition, association studies of the CA3 polymorphism with carcass traits in 140 $Yorkshire{\times}Meishan$ $F_2$ offspring showed that the NcoI PCR- RFLP genotype may be associated with variation in several carcass traits of interest for pig breeding. Allele B was associated with increases in lean meat percentage, loin eye height and loin eye area. Statistically significant association with backfat thickness was also found; pigs with the AB genotype had much less backfat thickness than AA or BB genotypes.

• 한국발생생물학회지:발생과생식
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• 제10권3호
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• pp.177-184
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• 2006

레티놀 결합 단백질(retinol-binding protein, RBP)은 고등 척추동물에서 혈류를 통해 특이적으로 레티놀을 표적세포에 운반해 주는 중요한 역할을 한다. 우리나라의 연안에 서식하고 있으며 산업적으로 중요한 조피볼락(Sebastes schlegeli)을 대상으로 4-nonylphenol(NP)가 RBP mRNA 발현에 미치는 영향을 구명하기 위해 간으로부터 cDNA library를 제작하고 RBP 단편의 염기서열을 분석하였다. 분석된 RBP mRNA 서열로부터 아미노산 서열을 추정하여 다른 종의 RBP 아미노산 서열과 비교한 결과 Sparus aurata와는 80%, 무지개 송어(Oncorhynchus mykiss)와는 72%, 유럽 뱀장어(Anguilla anguilla)와는 78%의 높은 상동성을 보였다. 조피볼락에서 4-NP에 대한 RBP와 VTG mRNAs 발현에 미치는 영향을 northern blot 분석 방법에 의해 조사하였다. 4-NP를 10 mg/kg BW로 주사한 실험구에서는 암수 모두 VTG mRNA 발현에 영향을 주지 않았으나, RBP mRNA 발현은 수컷에서 48시간 후에 감소하였다. 4-NP를 25 mg/kg BW으로 주사한 실험구에서는 24시간 후에 암수 모두 VTG mRNA 발현이 증가하였으며, RBP mRNA 발현은 48시간 후에 암수 모두 감소하였다. 이들 결과로부터 조피볼락에서는 4-NP 등의 에스트로겐 유사물질이 RBP와 VTG 유전자 발현에 상반되는 효과를 유도함을 알 수 있었다.