• 제목/요약/키워드: DnaB

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Cloning and Characterization of UV-B Inducible Chalcone Synthase from Grape Cell Suspension Culture System and Its Expression Compared with Stilbene Synthase

  • Song, Won-Yong;In, Jun-Gyo;Lim, Yong-Pyo;Park, Kwan-Sam
    • Journal of Photoscience
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    • 제7권2호
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    • pp.53-58
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    • 2000
  • We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

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효모재조합 DNA B형 간염백신의 면역효과에 관한 연구 (A Study of Immunogenicity and Reactogenicity of Hepatitis B Vaccine Made by Recombinant DNA Techniques in Yeast)

  • 민창홍;김교명;이규만
    • 대한미생물학회지
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    • 제21권2호
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    • pp.243-249
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    • 1986
  • A study of the immunogenicity and reactogenicity of two doses of lot H(10, 20 mcg), two doses of lot L (20, 40 mcg) of the Smith Kline-RIT recombinant DNA yeast-derived hepatitis B vaccine and a 20-mcg dose of the Merck Sharp and Dohme plasma-derived hepatitis B vaccine was conducted in young adults under randomized, double-blind conditions. Immunization was carried out according to a 0-, 1-, and 6-month vaccination schedule. Results indicated that the yeast-derived hepatitis B vaccine was well tolerated and immunogenic. Reactogenicity to both yeast- and plasma-derived vaccines was mild in severity and low in incidence with no significant differences appearing between the study groups. One month after the third dose, the yeast-derived vaccines induced a high degree of soroconversion ranging between 95.0% and 100%. The response was not lot or dose-dependent. The administration of the plasma-derived vaccine resulted in anti-HBs geometric mean titres statistically signifirantly higher than those elicited by the different yeast-derived hepatitis B vaccines one month after the third dose of vaccine but the difference was not large enough to be of great clinical significance.

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엽록체 DNA 염기서열 분석을 이용한 한국산 초롱꽃과 (Campanulaceae)의 계통유연관계 (Phylogenetic relationships of Korean campanulaceae based on chloroplast DNA sequences)

  • 김경아;유기억
    • 식물분류학회지
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    • 제42권4호
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    • pp.282-293
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    • 2012
  • 한국산 초롱꽃과 8속 28분류군과 외군 2분류군 등 총 30분류군에 대한 계통유연관계를 알아보기 위하여 엽록체 DNA의 atpB, atpB-rbcL, atpF-H, matK, rbcL, rpl16, rpoC1 그리고 trnL-F 지역의 염기서열 분석을 실시하였다. 8개 유전자 지역의 염기서열 자료를 융합하여 분석한 결과 도라지속과 더덕속이 가장 기부에 분계조를 형성하였고 애기도라지속과 초롱꽃속은 각각 독립적으로 유집되었다. 홍노도라지속과 영아자속은 잔대속-금강초롱꽃속 분계조를 위한 자매군을 형성하였으며, 금강초롱꽃속은 잔대속의 모시대절과 분계조를 형성하였고 잔대속은 크게 하나의 군을 형성하였다. 본 연구에서 다룬 엽록체 DNA 8개 지역의 염기서열 자료에 기초한 한국산 초롱꽃과의 계통분석 결과, 속 수준에서의 구분은 가능하였으나 절 또는 계열 등 잔대속 내에서의 일부 분류계급은 형태 형질에 의한 분류체계와 일치하지 않았다.

Gametophytic Abortion in Heterozygotes but Not in Homozygotes: Implied Chromosome Rearrangement during T-DNA Insertion at the ASF1 Locus in Arabidopsis

  • Min, Yunsook;Frost, Jennifer M.;Choi, Yeonhee
    • Molecules and Cells
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    • 제43권5호
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    • pp.448-458
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    • 2020
  • T-DNA insertional mutations in Arabidopsis genes have conferred huge benefits to the research community, greatly facilitating gene function analyses. However, the insertion process can cause chromosomal rearrangements. Here, we show an example of a likely rearrangement following T-DNA insertion in the Anti-Silencing Function 1B (ASF1B) gene locus on Arabidopsis chromosome 5, so that the phenotype was not relevant to the gene of interest, ASF1B. ASF1 is a histone H3/H4 chaperone involved in chromatin remodeling in the sporophyte and during reproduction. Plants that were homozygous for mutant alleles asf1a or asf1b were developmentally normal. However, following self-fertilization of double heterozygotes (ASF1A/asf1a ASF1B/asf1b, hereafter AaBb), defects were visible in both male and female gametes. Half of the AaBb and aaBb ovules displayed arrested embryo sacs with functional megaspore identity. Similarly, half of the AaBb and aaBb pollen grains showed centromere defects, resulting in pollen abortion at the bi-cellular stage of the male gametophyte. However, inheritance of the mutant allele in a given gamete did not solely determine the abortion phenotype. Introducing functional ASF1B failed to rescue the AaBb- and aaBb-mediated abortion, suggesting that heterozygosity in the ASF1B gene causes gametophytic defects, rather than the loss of ASF1. The presence of reproductive defects in heterozygous mutants but not in homozygotes, and the characteristic all-or-nothing pollen viability within tetrads, were both indicative of commonly-observed T-DNA-mediated translocation activity for this allele. Our observations reinforce the importance of complementation tests in assigning gene function using reverse genetics.

한국의 제주도에 서식하고 있는 노루(Capreolus pygargus tuanschanicus Satunin)의 미토콘드리아 DNA cytochrome b 유전자의 다양성 (Diversity of Mitochondrial DNA Cytochrome b Gene in Roe Deer (Capreolus pygargus tianschanicus Satunin) from Jejudo Island, Korea)

  • Koh, Hung-Sun;Yang, Beong-Guk;Yoo, Hye-Sook;Chun, Tae-Young
    • Animal Systematics, Evolution and Diversity
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    • 제16권2호
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    • pp.169-176
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    • 2000
  • 제주도산 노루 (C. Pygargus tianschanicus)의 분류학적 위치를 규명하기 위한 연구의 일환으로, 한국의 제주도에서 채집된 6마리의 노루 표본들을 이용하여 mtDNA의 cytochrome b 유전자의 부분적인 염기서열의 분석을 하였다. 밝혀진 세 haplotype간의 nucleotide Tamura & Nei's distance는 최대 0.005로써, 노루의 다른 아종 내의 다양성과 비슷한 정도였다. 또한 제주도산 노루의 cytochrome b 염기서열들과 GenBank에서 얻은 서 시베리아 지역의 노루인 C. p. Pygargus의 cytochrome b 염기서열간의 nucleotide distance는 평균 0.013였으며, C. p. tianschanicus는 65만년 전에 서 시베리아 지역의 노루인 C. p. Pygargus에서 분화되었을 것으로 판단된다.

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Ethanol Extract of Smilax glabra Induces Apoptotic Cell Death in Human YD10B Oral Squamous Cell Carcinoma Cells

  • Young Sun Hwang
    • 치위생과학회지
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    • 제23권3호
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    • pp.216-224
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    • 2023
  • Background: Smilax glabra has various pharmacological activities and is widely used as a herbal medicine. Although the incidence of oral cancer is low, the recurrence rate is high, and the 5-year survival rate is poor. It is necessary to search for anticancer drugs that increase the effect of cancer chemotherapy on heterogeneous oral tissues and reduce the side effects on normal cells. This study aimed to investigate the effects and mechanism of ethanol extract of Smilax glabra (EESG) as an anticancer drug for oral cancer. Methods: Smilax glabra root components extracted with 70% ethanol were used to analyze their effects on cancer cells. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay was performed for cytotoxicity analysis. Flow cytometry was performed to determine the cell cycle phase distribution. To observe apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling and γH2AX were detected by fluorescence microscope. The protein levels of cleaved PARP and caspase were analyzed using western blotting. The activation of procaspase-3 was confirmed by measuring caspase-3 activity. Results: EESG was no cytotoxic to normal gingival fibroblast but was high in YD10B oral squamous cell carcinoma (OSCC) cells. EESG treatment increased the subdiploid DNA content of YD10B cells by assessing DNA content distribution. Chromatin condensation and DNA strand breaks increased in YD10B cells treated with EESG. EESG-treated YD10B cells had high Annexin V and low propidium iodide levels, confirming that early apoptosis was induced. In addition, increased levels of γH2AX foci, a marker of DNA damage, were observed in the nuclei of EESG-treated YD10B cells. The EESG-treated YD10B cells also exhibited decreased procaspase-3 and procaspase-9 levels, increased PARP cleavage and caspase-3 activity. Conclusion: These results indicate that EESG inhibited cancer cell proliferation by inducing apoptosis in YD10B OSCC cells.

E. coli DNA 회복에 미치는 플라스미드 pKM101과 pSL4의 mutator 기능 (Mutator effects of plasmid pKM101 and pSL4 to E. coli DNA repair)

  • 전홍기;이상률;백형석
    • 미생물학회지
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    • 제28권2호
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    • pp.109-113
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    • 1990
  • Mutator 기능을 가지는 플라스미드 pKM101과 이의 돌연변이체 pSL4를 DNA 수복능력이 다른 Esherichia coli B/r(phr, recA, uvrA, uvrB)균주들에 도입하여 돌연변이원인 UV와 MNNG에 대한 보호효과와 mutagenecity를 조사형ㅅ다. pKM101과 pLS4의 mutator 기능과 보호효과는 repair 기능에 따라 다르나 전반적으로 두 플라스미드의 UV와 MNNG에 대한 저향성과 돌연변이율을 증가시켰고 pLS4는 pKM101보다 그효과가 높았다. 이러한 pLS4와 pKM101의 기능적 차이는 이들 플라스미드의 mutator 유전자상에 일어난 변이에 의한 것이라고 생각되었다.

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Anticarcinogenic and Antigenotoxic Effects of Bacillus polyfermenticus

  • Park, Eun-Ju;Kim, Kee-Tae;Kim, Cheon-Jei;Kim, Chang-Han;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.852-858
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    • 2004
  • The morbidity and mortality of colon cancer are increasing, because of the westernization of food habit. Probiotics such as lactic acid bacteria (LAB) have been known to play an important role in retarding colon carcinogenesis by possibly influencing metabolic, immunologic, and protective functions in the colon. In this study, we evaluated the effect of B. polyfermenticus SCD on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced DNA damage in CHO-K, cells and human lymphocytes, and on proliferation of human colon cancer cell. Using the Comet assay to detect DNA damage, we found that B. polyfermenticus SCD protected cells from the DNA damage induced by MNNG in $CHO-K_1$ cells and in human lymphocytes. B. polyfermenticus SCD was also found to inhibit the growth of colon cancer cells in a dose-dependent manner, detected by the MTT assay. These results indicate that B. polyfermenticus SCD has the potential to inhibit not only DNA damage induced by a carcinogen, but also the proliferation of colon cancer cells.

한국산 짱뚱어(Boleophthalmus pectinirostris)와 남방짱뚱어(Scartelaos gigas) (Gobiidae)의 분자유전학적 계통연관과 DNA 다형화 (Phylogenetic Relationship and DNA Polymorphism of Boleophthalmus pectinirostris and Scartelaos gigas (Teleostei: Gobiidae) of Korea)

  • 최기호;정의영;박갑만
    • 한국어류학회지
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    • 제25권3호
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    • pp.149-156
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    • 2013
  • 우리나라 망둑어과 어류 중 말뚝망둥어아과에 속하며 형태학적으로 흡사한 짱뚱어 (B. pectinirostris)와 남방짱뚱어(S. gigas)를 대상으로 12S rRNA, 16S rRNA 및 mitochondria cytochrome b 유전자의 염기서열 분석을 통해, 종내 및 종간의 분자유전학적 특성을 파악하고자 하였다. 짱뚱어 종내 개체간 유전적 차이를 밝히기 위해 mitochondria내에 3가지 유전자를 분석한 결과, 순천지역과 군산지역간의 염기서열은 12S rRNA는 총 434 bp (base pair)의 염기서열을 얻었으며 이를 비교 분석한 결과, 순천지역과 군산지역의 집단 간에는 100% 동일한 염기서열을 보였고, 16S rRNA 유전자는 총 484 bp의 염기서열 중 99.6%의 차이로 단지 2 bp가 치환된 것으로 나타났고, mitochondria cytochrome b 유전자의 경우, 444 bp의 염기서열을 얻었으며 두 지역 간 염기서열은 100%의 일치도를 보였다. 한국산 짱뚱어를 대상으로 한 동일한 종내에서 서식지 (서해, 남해)에 따른 유전적 차이나 지리적 거리의 차이가 있을 것으로 예상하였으나 이상의 3가지 유전자를 비교해 본 결과를 종합해 보면, 종내 개체 변이와 지역별 변이는 일어나지 않은 것으로 밝혀졌다. 또한 짱뚱어와 남방짱뚱어 2종간 비교 (inter-species)에서는 순천지역 짱뚱어와 해남지역 남방짱뚱어를 12S rRNA 유전자와 16S rRNA 유전자 및 mitochondria cytochrome b 유전자 염기서열을 비교 분석한 결과, 각각 96.1% (17 bp), 94.0 (29 bp), 그리고 82.9% (76 bp)로 나타나 두 종의 유전학적 차이는 크게 나타났다. 따라서 본 연구에서 나타난 짱뚱어와 남방짱뚱어 두 종간에 3.9~17.1%를 보여 서로 다른 종으로 볼 수 있을 것으로 사료되었다.

진딧물 rRNA 유전장에 특이적으로 결합하는 단백질 탐색 (Detection of the Specific DNA-binding Proteins for the Aphid rRNA)

  • O-Yu Kwon;Dong-Hee Lee;Tae-Young Kwon
    • 한국응용곤충학회지
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    • 제34권2호
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    • pp.100-105
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    • 1995
  • 정확한 in vitro 전사가 일어날 수 있는 진딧물의 세포추출액을 제조하였다. 전사를 직접 조절할 수 있는 단백질 인자를 규명하기 위하여 전사개시점과 그의 상류에 결합하는 DNA 결합단백질을 탐색했다. 전사개시점을 포함하는 단편 A(-194/23)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합했으며 전사개시점 상류의 DNA 단편 B(-393/-263)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합한 반면 DNA 단편 C(-263/-195)는 53kDa단백질만이 결합했다. 그리고 이들 DNA 결합단백질들의 DNA 결합 활성에는 양이온이 요구되었다.

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