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효모 유전자 발현용 (發現用) Promoter 개발에 관한 연구
정동효,정호권,박준희,심상국 ( Dong Hyo Chung,Ho Kwon Chung,Joon Hee Park,Sang Kook Shim ) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.1
The purpose of this study was the development of promoter for the lacZ` gene. Two heterologous promoter Ⅰ and Ⅱ of lacZ` gene were isolated from chromosomal DNA Bam HI fragment of yeast. The size of the promoter Ⅰ was estimated to be 2.5 kb and β-galacrosidase activity was 124.6 U/㎎ protein, and the size of the promoter Ⅱ was 4.0 kb and its β-galactosidase activity was 168.8 U/㎎ protein, respectively. The stability of the recombinant YEp plasmid in the transformant was from 52.7 to 67.4% at minimal medium. YIp plasmid was constructed from YEp plasmid, and expressed both in E. coli and yeast. The promoter Ⅰ aid Ⅱ iso-laced from yeast chromosomal DNA can be used for promoter of plasmid YEp and YIp.
온도조절형 발현 Vector 를 함유한 Phenylalanine 생산균의 분자육종
정동효(Dong Hyo Chung),심상국(Sang Kook Shim),이영춘(Young Chun Lee),정호권(Ho Kwon Chung) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.1
In order to produce phenylalanine without tyrosine co-production, we constructed various temperature-controllable expression vectors by insertion of lower expression of the tyrA gene into the plasmid pSY130-14. And tyrosine revertant to cultivate without addition of tyrosine, was selected from Escherichia coli strain AT2471[tyrA , thi ] by spontaneous mutation. The strain AT2471 harbouring plasmid pSY146A and the tyrosine revertant 5 harbouring plasmid pSY111-14 produced 12 g/ℓ and 15 g/ℓ of phenylalanine respectively in a 2.5ℓ jar fermenter at a constant temperature of 39℃ after 55 hours cultivation.
화학물질 분류 및 표시 국제기준의 국내 적용방안에 관한 화학물질 관련 업종별 실태 조사
황호순(Ho Soon Hwang),피영규(Young Gyu Phee),노영만(Young Man Roh),심상효(Sang Hyo Shim),김윤신(Yoon Shin Kim) 한국실내환경학회 2009 한국실내환경학회지 Vol.6 No.3
The main purpose of this research is to formulate best policy measure to minimize the expected problems when adopting and implementing GHS system in Korea and to draft the revision proposal for Industrial Safety and Health Act taking into account of unique domestic situation. The research method is to conduct a survey for 2 month period from early April to late May in 2006 to 830 randomly selected chemical manufacturing, importing and exporting, and consumption companies out of all the companies surveyed by the Ministry of Labor under 2004 Work Environment Status National Survey. A total of 610 survey was collected and analyzed. The results of this thesis is summarized as follows ; First, based on the survey analysis it is vital to conduct a national PR using pamphlet, internet, and daily newspaper and to provide technical assistance such as training expert and publishing GHS manual by expert organization such as Korea Occupational Safety and Health Agency (KOSHA) for early settlement of GHS in Korea. Second, it is also needed to give a grace period of 1 to 2 years to minimize the dramatic impact for industry, to encourage the establishment of the GHS team utilizing safety managers within companies, and to develop and distribute the standard GHS software by government. Third, taking into account of difficult situation of small companies, KOSHA needs to provide a full technical and financial support for companies with less than 100 employees and especially for chemical manufacturing companies. Fourth, it is also needed to operate an Interministrial GHS Committee (IGC) involving 7 related ministries for smooth GHS implementation and to develop an infra by sharing responsibilities among related ministries and establishing internationally recognized organization for hazard classification, labelling, and MSDS.
정동효,이기성,한면수,심상국 한국농화학회 1990 Applied Biological Chemistry (Appl Biol Chem) Vol.33 No.2
A bacterial strain which was capable of producing extracellular soymilk-clotting enzyme was isolated from soil samples during the course of screening test. The characteristics of the isolated strain K-324-7, indicated that the strain belonged to species of Bacillus cereus. The crude purification of this enzyme was precipitated by salting out with ammonium sulfate of 0.8 saturation. The optimal pH for the enzyme activity was at 6.l ∼7.0 and below 50℃. The optimal culture medium for the production of soymilk dotting enzyme were consisted of 0.2% glucose, 0.2% peptone, and 0.5% KH₂PO₄ with initial pH value of 6.5. The activity of enzyme was maximum when the microbe was cultured for 3 days at 35℃.
Bacillus 속이 생산하는 생전분 가수분해효소 유전자의 클로닝과 발현
정동효,심상국,김연계,김철호 中央大學校 遺傳工學硏究所 1989 遺傳工學硏究論集 Vol.2 No.1
생전분 분해 α-amylase를 강력하게 생산하는 Bacillus sp.를 분리하였다. E.coli-B. subtilis shuttle vector인 pYEJ001과 pHY300PLK를 사용하여 생전분 분해α-amylase 유전자를 E.coli와 Bacillus subtilis에 형질전환하였다. 클로닝된 α-amylase gene은 5.4kb HindⅢ 와 3.3kb EcoR V 단편의 크기였으며, E.coli내에서 이들 유전자는 비교적 안정하게 유지되었고 발현되었지만 Bacillus subtilis에서는 불안정하였으며 발현이 낮았다. 대장균내에서 생성된 생전분 분해 α-amylase는 그 효소의 생성량의 80%가 periplasmic space에 존재하였다. Bacillus sp which produces the raw starch digesting α-amylase screened. 5.4kb HindⅢ and EcoR V fragment of raw starch digesting α-amylase gene were cloned in the plasmid pYEJ001 or pHY300PLK which are E.coli-Bacillus subtilis vector using E.coli and Bacillus subtilis as a host,respectively. The cloned gene was certificated to hybridize to the Bacillus sp chromosomal DNA. The cloned gene was stably maintained and expressed in E.coli C 600,but was very unstable and low expressed in Bacillus subtilis 207-25. Analysis of α-amylase expressed in the transformants indicated that about 80% of α-amylase produced by the constructed plasmid of E.coli was localized in the periplasmic space and can be released by an cold osmatic shock.