• 제목/요약/키워드: gene integration

검색결과 321건 처리시간 0.026초

Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

  • Shin, Byung-Sik;Koo, Bon-Tag;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제4권4호
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    • pp.285-289
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    • 1994
  • An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The $lacl^q$ gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera: Arctiidae) and Plutella xylostella (Lepidoptera:Plutellidae).

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Gene Editing for Major Allergy Genes using Multiplex CRISPR-Cas9 System & Prime editing in Peanuts (Arachis hypogaea L.)

  • Min-cheol Kim;Tae-Hwan Jun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.194-194
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    • 2022
  • Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

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Gene Editing for Major Allergy Genes using Multiplex CRISPR-Cas9 System & Prime Editing in Peanuts (Arachis hypogaea L.)

  • Min-cheol Kim;Tae-Hwan Jun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.200-200
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    • 2022
  • Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

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에이전트 기반의 벼 기능 유전자 통합 데이터베이스 (An agent-based integrated database for rice functional genomics)

  • 이기열;신문수;안수영;정동훈;안진흥;정무영
    • 한국경영과학회:학술대회논문집
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    • 대한산업공학회/한국경영과학회 2006년도 춘계공동학술대회 논문집
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    • pp.1702-1706
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    • 2006
  • In the field of rice research, insertional mutants have become a valuable resource for studies of gene function. However, a well-designed database yet in the area of rice functional genomics. The relevant data are widely distributed and independently managed by the individual research groups. Heterogeneous data format in the distributed database systems causes many problems related to redundancy and compatibility. In this research, integration of the distributed databases using agent technology is pursued. In particular, a data integration agent, an ontology agent, a comparison agent, and resource agents are designed, whereby the integrated database is maintained. Moreover a framework for the web-based information system, which provides information to biologists and permits biologists to add new data to the database, is proposed. To establish an interoperable data format, an XML-based data model is also developed adopting ontology concept.

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Iron Chelator-Inducible Expression System for Escherichia coli

  • Lim, Jae-Myung;Hong, Mi-Ju;Kim, Seong-Hun;Oh, Doo-Byoung;Kang, Hyun-Ah;Kwon, Oh-Suk
    • Journal of Microbiology and Biotechnology
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    • 제18권8호
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    • pp.1357-1363
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    • 2008
  • The $P_{entC}$ promoter of the entCERA operon encoding enzymes for enterobactin biosynthesis in Escherichia coli is tightly regulated by the availability of iron in the culture medium. In iron-rich conditions, the $P_{entC}$ promoter activity is strongly repressed by the global transcription regulator Fur (ferric uptake regulator), which complexes with ferrous ions and binds to the Fur box 19-bp inverted repeat. In this study, we have constructed the expression vector pOS2 containing the $P_{entC}$ promoter and characterized its repression, induction, and modulation by quantifying the expression of the lacZ reporter gene encoding $\beta$-galactosidase. $\beta$-Galactosidase activities of E. coli transformants harboring pOS2-lacZ were highly induced in the presence of divalent metal ion chelators such as 2,2'-dipyridyl and EDTA, and were strongly repressed in the presence of excess iron. It was also shown that the basal level $\beta$-galactosidase expression by the $P_{entC}$ promoter was drastically decreased by incorporating the fur gene into the expression vector. Since the newly developed iron chelator-inducible expression system is efficient and cost-effective, it has wide applications in recombinant protein production.

A Unified Object Database for Biochemical Pathways

  • Jung, T.S.;Oh, J.S.;Jang, H.K.;Ahn, M.S.;Roh, D.H.;Cho, W.S.
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2005년도 BIOINFO 2005
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    • pp.383-387
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    • 2005
  • One of the most important issues in post-genome era is identifying functions of genes and understanding the interaction among them. Such interactions from complex biochemical pathways, which are very useful to understand the organism system. We present an integrated biochemical pathway database system with a set of software tools for reconstruction, visualization, and simulation of the pathways from the database. The novel features of the presented system include: (a) automatic integration of the heterogeneous biochemical pathway databases, (b) gene ontology for high quality of database in the integration and query (c) various biochemical simulations on the pathway database, (d) dynamic pathway reconstruction for the gene list or sequence data, (e) graphical tools which enable users to view the reconstructed pathways in a dynamic form, (f) importing/exporting SBML documents, a data exchange standard for systems biology.

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Stimulation of Actinorhodin Production by Streptomyces lividans with a Chromosomally-Integrated Antibiotic Regulatory Gene afsR2

  • Kim, Chang-Young;Park, Hyun-Joo;Yoon, Yeo-Joon;Kang, Han-Young;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제14권5호
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    • pp.1089-1092
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    • 2004
  • An actinorhodin nonproducing Streptomyces lividans was converted to an actinorhodin overproducer through a single chromosomal integration of an antibiotic regulatory gene, afsR2. This strain exhibited early actinorhodin production and an average of 37.5% higher productivity than the S. lividans containing multiple copies of afsR2 plasmid in a glucose-containing liquid culture.

Glucoamylase 유전자 STA의 염색체내 삽입에 의한 Saccharomyces diastaticus의 glucoamylase 생성능 향상 (Improvement of Glucoamylase Productivity of Saccharomyces diastaticus by Intergration of Glucoamylase Gene, STA, into Chromosomal DHA)

  • 안종석;맹준호;강대욱;황인규;민태익
    • 미생물학회지
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    • 제31권1호
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    • pp.48-53
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    • 1993
  • Saccharomyces distaticus 의 glucoamylase 생성능을 증진시킬 목적으로 STA1 유전자를 YIp vector 를 이용하여 염색체에 도입해 주고자 하였다. STA1 유전자 5.8-Kb 를 YIp vector 에 재조합하여 YIp-STA 를 재작하고, S. diastaticus GMT-11 (a, ura 3, STA1) 을 숙주균주로 하여 염색체의 STA1 유전자 부위에 homologous recombination 되어 삽입하도록 형질전환을 실시하였다. 이렇게 하여 glucoamylase 생성능이 모균주에 비해 최대 6배까지 증대된 다양한 형질전환체들을 얻을 수 있었다. 그리고 glycoamylase 생성능이 증대된 형질전환체들의 염색체 DNA 를 분리하여 Southern hybridization 을 실시한 결과 YIp-STA 가 multi-copy integration 되었음을 확인하였고, 또한 도입해 준 YIp-STA 는 세포분열인 30세대기간 동안 계속되었어도 안정하게 유지되었음을 알았다.

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Computational identification of significantly regulated metabolic reactions by integration of data on enzyme activity and gene expression

  • Nam, Ho-Jung;Ryu, Tae-Woo;Lee, Ki-Young;Kim, Sang-Woo;Lee, Do-Heon
    • BMB Reports
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    • 제41권8호
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    • pp.609-614
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    • 2008
  • The concentrations and catalytic activities of enzymes control metabolic rates. Previous studies have focused on enzyme concentrations because there are no genome-wide techniques used for the measurement of enzyme activity. We propose a method for evaluating the significance of enzyme activity by integrating metabolic network topologies and genome-wide microarray gene expression profiles. We quantified the enzymatic activity of reactions and report the 388 significant reactions in five perturbation datasets. For the 388 enzymatic reactions, we identified 70 that were significantly regulated (P-value < 0.001). Thirty-one of these reactions were part of anaerobic metabolism, 23 were part of low-pH aerobic metabolism, 8 were part of high-pH anaerobic metabolism, 3 were part of low-pH aerobic reactions, and 5 were part of high-pH anaerobic metabolism.