• Title/Summary/Keyword: Clone

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Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene of Bacillus stearothermophilus in Escherichia coli (Bacillus stearothermophilus Acetyl Xylan Esterase 유전자의 크로닝과 Escherichia coli에서의 발현)

  • 김인숙;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.542-548
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    • 1993
  • Bacillus stearothermophilus was shown to express multiple xylanolytic enzymes including acetyl xylan esterase. Genomic DNA of the strain partially digested with HindIII was ligated into the HindIII site of pBR322, and expressed in E. coli HB101 cells in order to clone the gene for acetyl xylan esterase. One transformant among 4000 screened formed a clear zone around its colony on the LB agar supplemented with 1.0% tributyrin. The functional clone harbored the recombinant plasmid pKMG5 with an insert of 5.1kb.

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Studies on the differentiation of Myoblasts: Molecular Cloning of differentiation related Genes in the Chick Embryonic Myoblasts by Differential Hybridization. (근세포 분화에 관한 연구: 차별 혼성화 스크리닝법에 의한 근원세포 분화 관련 유전자의 클로닝)

  • 강봉석;장세헌유병제양재섭
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.240-248
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    • 1994
  • 골격근 세포는 미분화 단핵 근원세포로부터 신장과 융합을 거쳐 다핵 횡문근섬유로 분화되어 가며 동시에 근특이 유전자의 발현이 선택적으로 일어난다. 본 연구에서는 계배 배양 근원세포의 분화동안 유전자 발현 조절 양상에 대한 연구를 위해, 계배 근원세포를 72시간 배양한 근섬유로부터 CDNA 라이브러리를 제작하였다. 이 cDNA 라이브러리를 미분화 단핵 근원세포(배양 36시간)와 분화된 다핵 근섬유(배양 72시간)의 poly(A)+ RNA 주형에서 합성된 [32P〕cDNA를 Probe로 사용한 differential plaque hybridization 방법으로 스크리닝하였다 분화된 다핵 근섬유 CDNA probe에 강한게 흔성화되는 CDNA clone을 선별하여 클로닝하였다. 선별한 CDNA clone 들 중 하나는 약 1.3 Kb 크기의 삽입절편을 갖고 있는 것으로 나타났고, 이 CDNA를 probe로 사용하여 northern blotting 한 결과, 이 CDNA엑 대한 유전자는 미분화 단핵 근원세포에서 분화된 다핵 근섬유로 분화가 진행됨에 따라 유전자 산물인 RNA 양이 증가되는 것으로 나타났다 또한 이 1.3 Kb CDNA에 대한 RNA의 크기는 약 2 7 Kb로 확인되었다.

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Isolation and Characterization of a CDNA Encoding a Protein Homologous to the Mouse 70 kDa Heat Shock Protein (생쥐 섬 유아세포에서 70 kDa 고온충격 단백질의 CDNA 클로닝과 염기서열 분석)

  • 김창환;정선미최준호
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.203-210
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    • 1992
  • Hsp70, a 70 kDa protein, is the maior protein expressed when cells are heat-shocked. A cDNA library from mouse ID13 cells was screened with the human hsp70 gene as a probe, and a positive clone was obtained. The positive clone was subcloned into puc19 and the precise restriction was obtained. The CDNA was sequenced by the Sanger's dideoxv termination method. Single open reading frame that codes for a protein of 70 kDa was found. The DNA sequence of the cloned mouse DNA shows great homology (66-90%) with other mouse hsp70 genes and somewhat less homology (50",) with E. coli hsp70 gene (dnak). With the exception of one amino acid, the protein sequence deduced from the CDNA is identical to the mouse that shock cognate protein 70 (hsc70) that is constitutivelv expressed at normal temperature. The result suggests that the cloned CDNA encodes a hsc70 family rather than a heatinducible family.mily.

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Differential Antitumor Activities of the Proteoglycans from the Mycelium of Lentinus Lepideus (잣버섯 균사체로부터 분리한 단백다당체의 암종에따른 선별적 항암작용)

  • Jin, Mi-Rim;Jung, Kyu-Sun;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.42 no.5
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    • pp.480-486
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    • 1998
  • Many antitumor and immune modulating components have been isolated from fungal extracts. In this study, the authors isolated the proteoglycans from cultured mycelia of Lentin us lepideus, including especially the acidic polysaccharide fraction, named lepidan. It was obtained by extraction with hot water followed by purification using DEAE cellulose anion exchange. To elucidate antitumor effects against different type of tumor, the proteoglycans were tested on sarcoma 180, C3H MCA clone 16 and P388 leukemia in vivo. Lepidan showed 58.3% of tumor inhibition against solid form of sarcoma 180 and 58.6% against MCA clone 16. But lepidan did not affect life span of mice against P388 leukemia. Also when Lepidan was applicated to MTT assay, it did not show any direct cytotoxicity against various tumor cells in vitro.

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A New Gene of Protein Related to Myoblast Fusion detected by Monoclonal antibidy (근원세포 융합과 관련된 새로운 유전자의 확인)

  • 박수정;이영주
    • The Korean Journal of Zoology
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    • v.38 no.1
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    • pp.49-54
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    • 1995
  • 본 연구자들은 근원세포를 면역시켜 얻은 hybidoma들을 검색하여. 계배 근원세포의 분화와 관련된 단백질을 인지하여 분화를 억제하는 대과가 있는 monoclonal antibody 3H35를 선별하여 그 항원을 확인한 바 있다(Kim et af.. (1992), Korean J. Zool 35 29-36) 본 연구에서는 λZAP에 cloning된 chicken muscle CDNA library들을 lacZ fusion protein으로 발현시켜 항체 3H35로 검색하여 그 유전자를 찾아내었다. 선별한 CDNA clone 중 C59의 삽입 절편은 1.6 kb이었고, 발현시킨 facE fusion protein 은 60 kDa로, f-galactosidase에 대한 항체에 반응하며 3H35와도 반응함을 immunoaffinitv adsorbant와 immunoblot으로 확인하였다 Clone C59의 삽입 절편의 염기서열을 분석한 결과, 실제 유전자는 1.6 kb 이상이며, 알려진 어느 다른 유전자와도 관련이 없는 새로운 근특이 유전자로 판단되었다. 아미노산으로 전환시켰을 때 31개의 특이한 서열이 7차례 반복된 부분이 나타났으며 이 서열의 23개가 일정하게 보존되어있고 나머지 서열의 아미노산의 polarity도 매우 유사하게 효존되어있다. 이들의 보존성이 극히 높은 것으로 보아 독특한 기능을 수행하는 domain으로 추정된다.

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Diversity of Microorganisms in Decaying Maize Stalks Revealed by a Molecular Method

  • Yang, Ming-Xia;Zhang, Han-Bo
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.367-370
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    • 2007
  • Microbial diversity in decaying maize stalk was characterized by constructing and analyzing rRNA gene clone library. Total 47 OTUs were obtained from 82 bacterial clones, including Proteobacteria (64.6%), Actinobacteria (30.5%), Bacteroidetes (2.4%) and Firmicutes (2.4%). Most proteobacterial clones were members of Rhizobium, Pseudomonas and Stenotrophomonas. Eighty-four percent of Actinobacteria was related to Microbacterium. Only 14 OTUs were identified from 124 fungal clones, including Ascomycota (88%) and Basidiomycota (12%). Sixty percent of Ascomycota were members of Eupenicillium and Paecilomyces but all Basidiomycota were close to Kurtzmanomyces nectairei.

Web Page Similarity based on Size and Frequency of Tokens (토큰 크기 및 출현 빈도에 기반한 웹 페이지 유사도)

  • Lee, Eun-Joo;Jung, Woo-Sung
    • Journal of Information Technology Services
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    • v.11 no.4
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    • pp.263-275
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    • 2012
  • It is becoming hard to maintain web applications because of high complexity and duplication of web pages. However, most of research about code clone is focusing on code hunks, and their target is limited to a specific language. Thus, we propose GSIM, a language-independent statistical approach to detect similar pages based on scarcity and frequency of customized tokens. The tokens, which can be obtained from pages splitted by a set of given separators, are defined as atomic elements for calculating similarity between two pages. In this paper, the domain definition for web applications and algorithms for collecting tokens, making matrics, calculating similarity are given. We also conducted experiments on open source codes for evaluation, with our GSIM tool. The results show the applicability of the proposed method and the effects of parameters such as threshold, toughness, length of tokens, on their quality and performance.

Molecular Characterization of Apolipophorin-III in the Fall Webworm, Hyphantria cunea Drury

  • Kim, Hong-Ja;Lee, Sang-Dae;Seo, Sook-Jae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.2
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    • pp.163-168
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    • 2001
  • We isolated and sequenced a cDNA clone corresponding to apolipophorin-III (apoLp-III) from the fall webworm, Hyphantria cunea. The cDNA for apoLp-III codes fer a 187-residue protein (561 bp) with a predicted molecular mass of 20 kDa. The calculated isoelectric point is 8.76. Multiple alignment analysis of the amino acid sequence revealed that H. cunea apoLp-III is most similar to that of Spodoptera litura (71.5% identity), followed by that of Manduca sexta (69.7% identity). They share five amphipathic $\alpha$-helices that are proposed to play a critical role in the binding of apoLp-III to lipophorin.

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Bombyx mori β-tubulin Promoter for High-level Expression of Heterologous Genes

  • Park, Kwanho;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.39 no.1
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    • pp.22-28
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    • 2019
  • We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.