• 제목/요약/키워드: Mass Identity

검색결과 242건 처리시간 0.034초

Saccharomyces cerevisiae에서 Neurospora crassa coq-4 유전자의 기능적 발현 (Functional Expression of the Neurospora crassa coq-4 Gene in Saccharomyces cerevisiae.)

  • 김은정;최상기;천재우;오계헌;이병욱
    • 미생물학회지
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    • 제40권2호
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    • pp.73-80
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    • 2004
  • Coenzyme Q는 지용성 퀴논 유도체로서 미토콘드리아의 내막과 원핵생명체의 세포막에 위치하는 전자전달계에서 전자 운반체로 이용되며 또한 항산화제의 기능도 갖는다. Coenzyme Q의 생합성에 관여하는 Saccharomyces cerevisiae의 coq4 유전자에 유사성을 나타내는 Neurospora crassa coq-4유전자를 클로닝하여 S. cerevisiae coq4 돌연변이체에서 기능적으로 발현하였다. 상보된 S. cerevisiae 균주들은 coenzyme $Q_{6}$의 생산능력을 회복하였으며 정상적인 성장률을 보였다. 또한 linoleic acid 및 linolenic acid와 같은 불포화지방산에 대한 낮은 감수성을 보였다. N. crassa의 COQ4 단백질은 39.7 kDa의 분자량을 갖는 347개의 아미노산으로 구성되어 있는 것으로 예상되며, S. cerevisiae의 Coq4p와 35%의 일치도 및 52%의 유사도를 보인다.

cDNA Cloning and mRNA Expression of A Cuticle Protein Gene Homo­logue from Protaetia brevitarsis

  • Kim Iksoo;Choi Yong Soo;Lee Eun Mee;Kim Mi Ae;Yun Enn Young;Ahn Mi Young;Jin Bynng Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제11권1호
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    • pp.67-70
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    • 2005
  • A cuticle protein gene, PbLCP12.1, from the white­spotted flower chafer, Protaetia brevitarsis, was isolated and characterized. The gene contains an ORF of 336 nucleotides capable of encoding a 113 amino acid polypeptide with a predicted molecular mass of 12,138 Da and pI of 4.15. The PbLCP12.1 protein contained a type-specific consensus sequence identifiable in other insect cuticle proteins. The deduced amino acid sequence of the PbLCP12.1 cDNA is most similar to Bombyx mori cuticle protein BmLCP18 (37$\%$ protein sequence identity). Northern blot analysis revealed that PbLCP12.1 showed the epidermis-specific expression.

A LIM Protein Gene Homologue of Protaetia brevitarsis: cDNA Cloning and mRNA Expression

  • Kim Iksoo;Choi Yong Soo;Lee Sun Young;Kim Mi Ae;Kim Seong Ryul;Hwang Jae Sam;Jin Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제11권1호
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    • pp.71-74
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    • 2005
  • A LIM protein gene homologue of the CRP (cysteine­rich protein) family in the whiter-spotted flower chafer, Protaetia brevitarsis, was cloned. The P. brevitarsis LIM protein cDNA encodes a 92 amino acid polypep­tide with a predicted molecular mass of 10,030 Da and a pI of 8.57. The P. brevitarsis LIM protein contains the cysteine-rich consensus sequence of LIM domain and the glycine-rich consensus sequence observed in the cysteine-rich protein family 1 (CRPl). The potential nuclear targeting signal is retained. The deduced amino acid sequence of the P. brevitarsis LIM protein cDNA showed 92$\%$ identity to another beetle, Apriona germari LIM protein. Northern blot analysis showed that P. brevitarsis LIM protein is highly expressed in epidermis and midgut, but not in the fat body.

Interactome Analysis of Yeast Glutathione Peroxidase 3

  • Lee, Phil-Young;Bae, Kwang-Hee;Kho, Chang-Won;Kang, Sung-Hyun;Lee, Do-Hee;Cho, Sa-Yeon;Kang, Seong-Man;Lee, Sang-Chul;Park, Byoung-Chul;Park, Sung-Goo
    • Journal of Microbiology and Biotechnology
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    • 제18권8호
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    • pp.1364-1367
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    • 2008
  • Oxidative stress damages all cellular constituents, and therefore, cell has to possess various defense mechanisms to cope. Saccharomyces cerevisiae, widely used as a model organism for studying cellular responses to oxidative stress, contains three glutathione peroxidase (Gpx) proteins. Among them, Gpx3 plays a major defense role against oxidative stress in S. cerevisiae. In this study, in order to identity the new interaction proteins of Gpx3, we carried out two-dimensional gel electrophoresis after immunoprecipitation (IP-2DE), and MALDI-TOF mass spectrometry. The results showed that several proteins including protein disulfide isomerase, glutaredoxin 2, and SSY protein 3 specifically interact with Gpx3. These findings led us to suggest the possibility that Gpx3, known as a redox sensor and ROS scavenger, has another functional role by interacting with several proteins with various cellular functions.

Optimal Conditions for the Post-Harvest Storage of Rhizoids of the Brown Seaweed Undaria pinnatifida (Phaeophyta) for Arachidonic Acid Production

  • Khan, Mohammed Nurul Absar;Kang, Ji-Young;Park, Nam-Gyu;Choi, Jae-Suk;Cho, In-Soon;Hong, Yong-Ki
    • Fisheries and Aquatic Sciences
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    • 제15권2호
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    • pp.163-168
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    • 2012
  • The non-utilized biomass of the aquacultured seaweed Undaria pinnatifida, particularly the rhizoid, is an alternative source of arachidonic acid (AA). Of the five aquacultured kelps that were tested, U. pinnatifida yielded the highest amount of AA, which was isolated from the rhizoids. Its identity (C20:4 n-6) was confirmed from gas chromatography-mass spectrometry spectral data. The optimal conditions for post-harvest storage or pretreatment of the rhizoids in Provasoli's enriched seawater for AA extraction were determined to be pH 7.8, 2% $CO_2$-enriched air, 20 ${\mu}mol\;m^{-2}\;s^{-1}$ light, and $10^{\circ}C$. Under these conditions, the AA content after 1 day of storage was enhanced by up to 127%. In the absence of light under ambient aeration, the AA content after 1 day of storage diminished to 90%. Rhizoids collected late in the season (April and May) contained the highest amounts of AA (approximately 2.5 mg/g tissue).

Cloning and Characterization of ${\alpha}-Glucosidase$ Gene from Thermophilic Bacillus sp. DG0303

  • Lee, Yong-Eok
    • Journal of Microbiology and Biotechnology
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    • 제10권2호
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    • pp.244-250
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    • 2000
  • An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

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Expression and In Vitro Activity of Recombinant Canstatin in Stably Transformed Bombyx mori Cells

  • Lee, Ji-Hye;Lee, Jong-Min;Jeon, Hwang-Bo;Shon, Bong-Hee;Yang, Jai-Myung;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
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    • 제19권7호
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    • pp.685-689
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    • 2009
  • We describe the expression of recombinant canstatin from stably transformed Bombyx mori BmS (BmS) cells. Recombinant canstatin was secreted into a culture medium with a molecular mass of approximately 29 kDa. Densitometric scanning showed that the secreted canstatin accounted for approximately 91% of the total canstatin production. Recombinant canstatin was also purified to homogeneity using a simple one-step Ni-NTA affinity fractionation. The identity of the purified protein was confirmed as human canstatin by nano-LC-MS/MS analysis. Purified recombinant canstatin inhibited human endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition ($ED_{50}$) for recombinant canstatin expressed in stably transformed BmS cells was approximately 0.64 ${\mu}g/ml$. A maximum production level of 11 mg/l recombinant canstatin was obtained in a T-flask culture of BmS cells after 6 days of incubation.

Cloning and Characterization of the Cu,Zn Superoxide Dismutase (SOD1) cDNA from the Spider, Araneus ventricosus

  • Choi Young Soo;Choo Young Moo;Li Jianhong;Sohn Hung Dae;Jin Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권1호
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    • pp.73-77
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    • 2005
  • A Cu,Zn superoxide dismutase (SOD1) cDNA was cloned from the spider, Araneus ventricosus. The A. ventricosus SOD1 (AvSOD1) cDNA contains an open reading frame of 495 bp encoding 165 amino acid polypeptide with a predicted molecular mass of 17,114 Da and pI of 6.55, and possesses the typical metal binding ligands of six histidines and one aspartic acid common to SOD1s. The deduced amino acid sequence of the AvSOD1 cDNA showed $51\%$ identity to Ceratitis capitata SOD1, and $50\%$ to SOD1 sequences of both Drosophila melanogaster and Chymomyza amoena. Northern blot analysis revealed the presence of AvSOD1 transcripts in all tissues examined.

A Cloning of Novel Esterase from a Metagenomic Library

  • Yoon, Sang-Young;Kim, Seung-Bum;Ryu, Yeon-Woo
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
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    • pp.243-246
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    • 2005
  • A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

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21세기 패션에 나타난 인트라컬추럴리즘(Intraculturalism)에 관한 연구 (A Study on Intraculturalism in 21st Century Fashion)

  • 양숙희;정세희
    • 복식
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    • 제59권1호
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    • pp.119-135
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    • 2009
  • The purpose of this study was to examine formative aesthetic characteristics and aesthetical value of the intraculturalism expressed in contemporary fashion and to confirm the functions of intraculturalism to establish, visualize, perform the racially indeterminate, ethnically neutral, culturally diverse or ambiguous identity. For this study, the applications of the intraculturalism shown in mass media and consumer culture, such as music, fashion advertisements and collections of high fashion designers from 2004 to 2008 have been analyzed and compared. The results were as follows: The Intraculturalism is reflected in the muticultural music such as Afropean, Jawaiian, Reggaeton and Asian Hip Hop. Intracultural music genres create the hybrid music and fashion culture through mixing, matching and blending one and another culture. Advertisement campaigns for Louis Vuitton, YSL Beauty, Gap and H&M stores have all purposely highlighted models with mixed racial heritage. It is reflected in the latest youth fashion market trend using face that are ethnically ambiguous. The increasingly multiracial, multicultural population is due to intermarriage and waves of immigration. The rising mixed race designers, Narciso Rodriguez, Hussein Chalayan, Vera Wang and DooRi Chung, not only compromise and amalgamate different cultural elements of their heritage and contemporary life but also create new look and fashion image. The characteristics of intraculturalism expressed in the 21st century fashion could categorized into de-genre, de-nationality, de-race and de-culture.