• Title/Summary/Keyword: 재조합

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The Dependency of the Expression Level of Recombinant Protein by the Drop of Alkali Consumption Rate after Induction (발현유도에 의한 알칼리 소비속도의 감소가 재조합 단백질 생산에 미치는 영향)

  • Hur, Won
    • KSBB Journal
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    • v.21 no.4
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    • pp.236-240
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    • 2006
  • IPTG induction caused a sudden drop of alkali consumption rate during cultivation of a recombinant E. coli with ${\beta}$-galactosidase structural gene under T7 promoter on a plasmid. A series of batch cultivations showed the positive correlation of the decrease of alkali consumption and the level of expression. However, repeated IPTG induction did not cause any variation of alkali consumption rate. Supplementation of medium even at stationary phase enhanced the level of ${\beta}$-galactosidase expression. These results suggests that the drop of alkali consumption rate by IPTG induction represents the rate of expression.

Occurrence of Tn3 Sequence Upstream of aacC2 Gene in Gentamicin Resistance R Plasmids (Gentamicin 저항성 R 플라스미드에 존재하는 aacC2 유전자의 상류부위에서 Tn3의 출현)

  • 한효심;김남덕;이영종;이효연;정재성
    • Korean Journal of Microbiology
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    • v.33 no.3
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    • pp.165-169
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    • 1997
  • Two gentamicin resistance R plasmids, pGM5 and pGM6, containing aacC2 gene were selected from environmental isolates. The gentamicin resistance determinants of R plasmids were cloned into the BamHI site of pUC18. Restriction enzyme map of inserted region of recombinant plasmids, pSYS and pSY6, and PCR results indicated that Tn3 sequence was located upstream of gentamicin resistance gene. Based on the restriction maps and susceptibility tests, it was concluded that the sequence of bla and 3' inverted repeat of Tn3 play a important roles in the expression of gentamicin resistance gene.

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Improved Recombinant ,$\beta$-Galactosidase Production Using Medium Additives at AcNPV Infection of Insect Cells in Batch and Continuous Two-Stage Bioreactors (회분식과 연속식 2단계 생물반응기에서 AcNPV의 곤충세포에의 감염시 배지 첨가물을 이용한 재조합 $\beta$-Galactosidase 생산의 증진)

  • 김지선;이기웅
    • KSBB Journal
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    • v.9 no.3
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    • pp.294-298
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    • 1994
  • The medium additives such as CaCl2, glucose, fructose, glutamine, glutamate and lipids were examined to enhance recombinant ${\beta}$-galactosldase(${\beta}$-gal) production in batch and continuous two-stage bioreactor systems. The presence of each medium additive such as CaCl2, fructose, glutamate, cholesterol and tocopherol at AcNPV infection of Sf 21 cells had an effect on improved ${\beta}$-gal production. The recombinant ${\beta}$-gal production using the infection media supplemented with a mixture of 30mM $CaCl_2$, 2.2mM fructose, 4.1mM glutamate and 0.34mM cholesterol was increased by about 40%.

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Expression of recombinant plasmids harboring glucoamylase gene STA in saccharomyces cerevisiae (Glucoamylase 유전자 STA를 포함한 재조합 플라스미드들의 saccharomyces cerevisiae에서의 발현)

  • 박장서;박용준;이영호;강현삼;백운화
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.181-187
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    • 1990
  • STA gene coding glucoamylase was introduced into haploid Saccharomyces cerevisiae SHY3 and polyploid Saccharomyces cerevisiae 54. We constructed the recombinant plasmid by substituting the promoter region of alcohol dehydrogenase isoenzyme I gene for that of STA gene to increase the expression of STA gene and found that the activity of glucoamylase was increased in transformants. The plasmid stability was improved remarkably when we got the STA gene into the plasmid which had centromere. The activity of glucoamylase and transformation frequency of it, however, was decreased because of low copy number. Industrial polyploid strain was transformed with the recombinant plasmid having the $2\mu$ origin of replication and STA gene. It produced more alcohol than host when fermented in liquefied starch media. The industrial strain, however, was not transformed with the autonomously replicating plasmid containing centromere.

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재조합 인간 GM-CSF의 수용체에 관한 연구

  • 이부연;최상운;이정옥;공재양
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.307-307
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    • 1994
  • 본 실험에서는 유전자 재조합으로 제조한 〔$^{125}$/I〕-labeled human GM-CSF를 사용하여 GM-CSF의 HL-60 cell의 표면에 존재하는 GM-CSF 수용체의 특성을 밝히고 수용체에 대한 binding parameter를 확인하고 Immunex(미국) 사에서 제조한 Prokine(Sargramostim)과 Sigma사에서 구입한 GM-CSF(C-9666)를 표준물질로 하여 (주) Lucky에서 제조한 GM-CSF(LBD-005)의 수용체에 대한 결합율을 측정, 각각 비교하고자 하였다. 한편 LBD-005는 glycosylation된 form과 안된 form의 혼합물이므로 당화의 정도가 수용체에 대한 결합에 미치는 영향을 알아보기 위해 glycosylated form과 혼합물(LBD-005)의 수용체에 대한 결합율을 측정 비교하였다.

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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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