• Title/Summary/Keyword: -galactosidase

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유전공학기법을 이용한 새로운 당뇨병 치료제의 개발 연구

  • 이승엽;이추희;남두현
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.138-138
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    • 1993
  • 인슈린 유사체를 유전공학적 방법으로 생산하여 새로운 당뇨병 치료제로써의 가능성을 타진한다. 인슈린 B 사슬의 C 말단 아미노산 codon을 threonine 대신을 methionine을 지령하도록 하고 여기에 인슈린 A사슬을 지령하는 염기를 바로 부착시켜, 이를 대장균에서 발현시키므로써 외사슬 인슈린 선구체를 제조하고, 이를 분리 정제한 다음 취화브롬 으로 절단하므로서 ($B^{30}$-homoserine) 인슈린을 제조한다. 1. 외사슬 인슈린 전구체 유전자를 합성한 후 대장균의 발현 운반체내 e-galactosidase 유전자와 융합시켜 도입하므로서 재조합된 융합단백질을 생산하였다. 2. 재조합 대장균을 발효한 후 urea 융합단백질을 분리하고 DEAE-Sephacel과 Sephadex G-200을 이용하여 순수 정제하였다.

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Mutagenic Effect of Steroidal Saponins from Smilax china Rhizomes (토복령(Smilax china)의 Steroid Saponin이 돌연변이원성에 미치는 영향)

  • Kim, Sung-Whan;Son, Kun-Ho;Chung, Kyu-Charn
    • YAKHAK HOEJI
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    • v.33 no.5
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    • pp.285-289
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    • 1989
  • Pontential mutagenic and antimutagenic activities of four steroidal saponins from Smilax china rhizomes were investigated. These saponins did not revealed mutagneicity in the Ames and SOS umu test. For antimutagenic activity by SOS umu test, two spirostanol glycosides, dioscin and gracillin, inhibited the activity of ${\beta}-galactosidase$ induced by AF-2, but their proto-type furostanol glycosides did not show this activity.

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Carbon Source-Dependent Regulation of the Schizosaccharomyces pombe pbh1 Gene

  • Kim, Su-Jung;Cho, Nam-Chul;Ryu, In-Wang;Kim, Kyung-Hoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.689-693
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    • 2006
  • Pbh1, from the fission yeast Schizosaccharomyces pombe, is a baculoviral inhibitor of apoptosis (IAP) repeat (BIR) domain-containing protein. Its unique encoding gene was previously found to be regulated by nitric oxide and nitrogen starvation. In the current work, the Pbh1-lacZ fusion gene was used to elucidate the transcriptional regulation of the pbh1 gene under various carbon sources. When fermentable carbon sources, such as glucose (at a low concentration of 0.2 %), sucrose (2.0 %) and lactose (2.0 %), were the sole carbon source, the synthesis of $\beta$-galactosidase from the Pbh1-lacZ fusion gene was reasonably enhanced. However, the induction by these fermentable carbon sources was abolished in the Pap1-negative S. pombe cells, implying that this type of induction of the pbh1 gene is mediated by Pap1. Ethanol (2.0%), a nonfermentable carbon source, was also able to enhance the synthesis of $\beta$-galactosidase from the fusion gene in wild-type cells but not in Pap1-negative cells. The results indicate that the S. pombe pbh1 gene is up-regulated under metabolic oxidative stress in a Pap1-dependent manner.

Inhibition of Ethylene Action Related to Poststorage Softening by 1-Methylcyclopropene Treatment in 'Fuyu' Persimmon Fruits (1-Methylcyclopropene 처리에 의한 '부유' 단감 과실의 저장 후 연회 현상과 관련된 에틸렌 작용의 억제)

  • Kim, Mi-Ae;Ahn, Gwang-Hwan;Lee, Seung Koo;Choi, Seong-Jin
    • Horticultural Science & Technology
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    • v.19 no.4
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    • pp.545-549
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    • 2001
  • The relationship between ethylene action and flesh softening in post-storage persimmon fruits was investigated by treating the fruits with 1-MCP. The patterns of firmness change caused by various treatments with 1-MCP and ethylene were similar to those of changes in cell wall-degrading enzyme activities, including cellulase, PG, PME, and ${\beta}$-galactosidase. Moreover, the activities of these enzymes were inhibited by 1-MCP treatment. These results show that the cell wall-degrading enzymes influenced by ethylene account for the flesh softening process of post-storage persimmon fruit. The ethylene production of fruits, as measured by ACC content and ACC oxidase activity, nevertheless, was not influenced by 1-MCP treatments. It is suggested that the flesh softening phenomena in post-storage persimmon fruits is correlated to the ethylene responsiveness of tissue rather than the ethylene production rate per se.

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Direct Analysis of the Transcription of Escherichia coli rnpB Gene Harbored in a Multicopy Plasmid during Bacterial Growth

  • Park, Jeong-Won;Jung, Young-Hwan;Park, Bo-Hyun;Jeoung, Yeon-Hee;Lee, Young-Hoon
    • BMB Reports
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    • v.29 no.3
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    • pp.221-224
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    • 1996
  • To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

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Construction of New Transfer Vector of Nuclear Polyhedrosis Virus of the Silkworm, Bombyx mori (누에 핵다각체병 바이러스를 이용한 새로운 전이 벡터의 제작)

  • 우수동;김우진;진병래;강석권
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.46-51
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    • 1995
  • In order to develope baculovirus expression vector system, we constructed new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. The promoter region containing only adenine of translation start codon of polyhedrin gene was cloned by polymerase chain reaction technique. And the 5' and 3' leader regions of polyhedrin gene was sequentially cloned. The polyhedrin coding gene was deleted from the +2 to the +597 position. As the result, we constructed new transfer vector which has EcoRI, SacI and KpnI sites for the cloning sites of foreign gene. New transfer vector was named as pBmKSKl. Escherichia coli $\beta$-galactosidase gene as foreign gene was inserted into pBmKSKl, under the control of the polyhedrin promoter and expressed in B. mori cells. The result showed that the new transfer vector pBmKSK1 is functional.

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Temperature-Dependent Expression of Escherichia coli Thioredoxin Gene

  • Lee, Jin-Joo;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • BMB Reports
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    • v.33 no.2
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    • pp.166-171
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    • 2000
  • Thioredoxin is a multifunctional protein that is ubiquitous in microorganisms, animals and plants. Previously, the expression of the Escherichia coli thioredoxin gene (trxA) was found to be negatively regulated by cAMP. In the present study, the effect of temperature on the expression of the E. coli trxA gene was investigated. In order to examine the temperature effect, the fusion plasmid pCL70 that harbors the E. coli trxA P1P2 promoter was used. The other two fusion plasmids, pJH3 and pMH521 that were constructed in different vectors which harbor the E. coli trxA P2 promoter, were also used. When the E. coli strain MC1061/pCL70 was grown in a rich medium at $25^{\circ}C$, $34^{\circ}C$ and $42^{\circ}C$, the cells grown at $42^{\circ}C$ gave the highest $\beta$-galactosidase activity. The E. coli MC1061/pJH3 and MC1061/pMG521 cells showed increased $\beta$-galactosidase activity after the shift of the culture temperature to $42^{\circ}C$. The wild-type trxA gene of the E. coli MC1061 cells produced much higher thioredoxin activity at the higher temperature. These results support the conclusion that the E. coli trxA gene is regulated in a temperature-dependent manner. Especially the expression from its P2 promoter appeared to be sensitive to temperature.

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