• 한국식품영양과학회지
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• 제30권5호
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• pp.802-805
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• 2001

산업적으로 lysine 발효 산업에 이용되고 있는 B. lactofermentum으로부터 lysine 생합성에 관여하는 tetrahyrodipicolinate N-succinyl transferase를 지령하는 dapD 유전자를 E. coli의 dapD 결손변이주와의 complementation test를 통하여 cloning하였다. 재조합 plamid는 3.6 kb의 DNA 단편을 함유하고 있었으며 Southern blot hybridization을 통하여 dapD 유전자는 B. lactofermentum으로부터 유래하였으며 염색체 DNA내에 single copy로 존재함을 알 수 있었다. 또한 lysine 생성량 분석을 통하여 E. coli에서 B. lactofermentum dapD 유전자의 발현을 확인하였다.

• Animal Systematics, Evolution and Diversity
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• 제37권4호
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• pp.354-357
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• 2021

In Korean waters, the cirolanid isopod, Eurydice longiantennata Nunomura and Ikehara, 1985 has been reported only from the subtidal zone of Jeju island. We obtained the mitochondrial cytochrome c oxidase subunit I (COI) sequences of this species and determined the DNA barcoding data of E. longiantennata based on a genetic comparison of E. longiantennata and its congeners. The intra-specific genetic distance between the three COI sequences of E. longiantennata ranged from 0 to 0.6%. The inter-specific distances between E. longiantennata and other cirolanid isopods ranged from 24 to 33.2%. In this study, we provided the DNA information of E. longiantennata with a morphological diagnosis and images of the species.

• 한국양식학회지
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• 제11권4호
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• pp.557-564
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• 1998

무지개송어의 초대배양 간세포에 있어서 Vitellogenin (VTG)의 합성 유도는 전기영동적 수법에 의해 행하였다. 간세포는 7일 동안 phositively charged dish를 이용한 배양으로 단층 확산되었다. 배양 7일의 간세포 생존율은 $E_2$의 유무에 따라 각각 20.7% 및 23.6%가 감소하였으며, DNA함유량도 각각 13.7% 및 14.0%가 감소하였다. $E_2$ 첨가군과 대조군간에는 생존율과 DNA 함유량에 있어서는 유의한 차이가 나타나지 않았다. 그리고, 총 단백질에 대한 VTG의 비율은 $E_2$$10^{-6}$ M의 첨가시에 최대치를 나타내었다. 그러나, 고농도($10^{-5}$M)에서는 오히려 감소하는 경향을 나타내었다.

• 수산해양기술연구
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• 제58권1호
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• pp.39-48
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• 2022

Using environmental DNA (eDNA) in the fisheries and oceanography fields, research on the diversity of biological species, the presence or absence of specific species and quantitative evaluation of species has considerably been performed. Up to date, no study on eDNA has been tried in the area of fisheries acoustics in Korea. In this study, the biomass of a dominant species in the northwestern waters of Jeju Island was examined using 1) the catch ratio of the species from trawl survey results and 2) the ranking ratio of the species from the eDNA results. The dominant species was Zoarces gillii, and its trawl catch ratio was 68.2% and its eDNA ratio was 81.3%. The Zoarces gillii biomass from the two methods was 7199.4 tons (trawl) and 8584.6 tons (eDNA), respectively. The mean and standard deviation of the acoustic backscattering strength values (120 kHz) from the entire survey area were 135.5 and 157.7 m²/nm², respectively. The strongest echo signal occurred at latitude 34° and longitude 126°15' (northwest of Jeju Island). High echo signals were observed in a specific oceanographic feature (salinity range of 32-33 psu and the water temperature range of 19-20℃). This study was a pilot study on evaluating quantitatively aquatic resources by applying the eDNA technique into acoustic-trawl survey method. Points to be considered for high-quality quantitative estimation using the eDNA to fisheries acosutics were discussed.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.800-807
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• 2022

Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.

• 생명과학회지
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• 제13권4호
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• pp.541-550
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• 2003

본 실험에서는 C형 간염바이러스 (HCV)의 외피 단백질인 E2 당단백질에 결합하는 세포단백질들을 클로닝하기 위해 간세포 cDNA를 phage 표면에 발현시킨 phage library를 제작하였고, 12-mer peptide library와 함께 E2 단백질에 대해 panning을 실시하였다. 검색결과 세포내 신호전달과 cytoskeleton 구성에 관여하는 tensin, membrane protein band 4.1 등 세포질내 단백질과 CCR7, CKR-L2, insulin-like growth factor-1 receptor 등 세포막 단백질 등이 확인되었다. 이들 단백질들을 발현하는 phage들은 수용성 E2단백질을 이용한 결합중화반응 결과 E2 단백질에 특이적으로 결합함이 확인되었다. 사람 T 세포에서 주로 발현되는 CCR7 유전자를 PHA로 활성화된 사람 T 세포의 total RNA를 이용하여 증폭하고 클로닝하였다. 293T 세포에 transfection시켜 단백질 발현양상을 flow cytometer로 분석하여 70% 이상의 세포들이 CCR7을 발현하고 있음을 관찰하였다. 수용성 E2 단백질을 CCR7이 transfection된 세포와 mock transfection된 대조군 세포에 각각 반응시킨 결과 dose-dependent 양상으로 CCR7에 결합하였다.

• 한국연초학회지
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• 제23권2호
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• pp.103-108
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• 2001

Hog cholera virus(HCV) was purified from virus infected Bovine kidney cells. From this virus, total protein was analyzed by SDS-PAGE gel electrophoresis and about 55 kDa band of E2 envelope protein was detected. The viral RNA was purified and E2 cDNA was amplified by RT-PCR. E2 cDNA fragment was cloned to PCRII-TOPO cloning vector and named pE2. The analysis of nucleotide sequence showed that this E2 cDNA fragment inserted into pE2 was 1191 nucleotides long and coded 397 amino acids.

• Nutritional Sciences
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• 제8권4호
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• pp.262-267
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• 2005

Reactive oxygen species can be generated in the skin by hair dyeing. The aim of this study was to find out the effects of the oxidative-type hair dye application in young women on the antioxidant systems. We investigated the lipid peroxide levels, glutathione (GSH) levels, and the antioxidant enzyme activities including superoxide dismutase (SOD), glutathione peroxidase (GSHPx) in plasma and erythrocytes and catalase (CAT) in erythrocytes, and DNA damages in lymphocytes. Also, plasma concentrations of antioxidant vitamins, vitamin A and E, were measured and the correlations between various antioxidant parameters and oxidative damages were evaluated The antioxidant enzyme activities in plasma (GSHPx) and in erythrocytes (SOD and CAT) were decreased significantly after hair dyeing. 1be lipid peroxide and GSH levels were not affected in both plasma and erythrocytes. No significant difference was found in the concentrations of both vitamin A and E between before and after hair dyeing. However, DNA damages expressed as the tail extent moment (TEM) and tail length (TL) were significantly (p<0.001) increased. The plasma vitamin E concentration was correlated with DNA damages (TEM: r=-0.590, p<0.01 and TL: r=-0.533. p<0.01) and RBC SOD activity (r=0.570, p<0.05). In turn, RBC SOD activity was significantly correlated with both plasma MDA levels (r=-0.412, p<0.05) and DNA damages (TM: r=-0.546, p<0.01, TL: r=-0.493, p<0.01). Our results demonstrated that the exposure to hair dyeing produced lymphocyte DNA damage and modification of the antioxidant enzyme activities. Also, there were very strong associations between plasma vitamin E concentration, RBC SOD activity and DNA damage induced by hair dyeing. It suggests that the antioxidant status of a subject is likely to be related to the extent of the harmful effects caused by hair dyeing.

• Animal cells and systems
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• 제3권2호
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

• Journal of Ginseng Research
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• 제13권1호
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• pp.37-41
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• 1989

Benzo(a)pyrene(BP)의 monooxygenase(AHH)에 의해서 생성된 반응성 대사물질들의 in vitro DNA와의 결합 및 BP 대사에 관여하는 효소들의 활성도에 미치는 인삼 물추출물의 영향을 조사하였으며, DNA-BP metabolite adduct들은 Sephadex LH-20 column으로 chromatography하여 5개의 major peak 들을 얻었다. 이 peak 들을 극성이 큰 순서대로 A부터 E까지 임의로 정하고 5개의 peak들을 7,8-diol-9,10-oxide(A), 7,8-oxide(B). 4,5-oxide(C), 9-HO-BP(D & E) adduct들로 잠정적으로 확인하였다. Peak A, C, D 그리고 E는 각각 대조군의 30, 15, 20 그리고 30%로 감소되었으며 peak B는 의미있는 변화를 보이지 않았다. DNA-BP 결합 억제와 관련하여 in vitro와 in vivo 투여시의 경향이 유사하여 EH의 활성도만 BP투여 대조군보다 38%정도 의미있게 유도되었다.