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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.
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정동효,윤광로,박준희 中央大學校 食糧資源硏究所 1989 食糧資源硏究所 論文集 Vol.1 No.1
Caprylic acid, caproic acid, lauric acid, myristic acid, palmitic acid등 의 포화지방산과 ascorbic acid로 에스테르를 합성하여 이들 에스테르의 순도 확인한 후 항균력 실험과 여러 가지 미생물에 대한 최소 증식저지농도와 식품저장의 보존제로서 응용 실험을 하여 다음과 같은 결과를 얻었다. GC/MS, IR,NMR,TLC 결과를 종합할 때 ascorbyl-6-caprylate,ascorbyl-6-caprate, ascorbyl-6-laurate, ascorbyl-6-myristate, ascorbyl-6-palmitate의 5종이 순도가 만족할만한 에스테르로 합성되었고 ascorbic acid의 6번째 탄소 위치에 지방산이 에스테르화되는 것을 확인하였다. 또 이들 에스테르들의 항균력은 그람양성 세균이 그람음성 세균보다 항균활성이 높았다. 그리고 에스테르들 중에서 ascorby-6-laurate가 다른 에스테르들보다 강한 항균활성이 있었다. Ascorbyl fatty acid ester는 그람양성 세균인 Bacillus subtilis, Staphylococcus aureus, Lacctobacillus brevis등에 대하여 항균활성이 높았고, 그람음성 세균인 Escherichia coli에서는 500㎍/ml농도에서도 항균활성이 없었다. 그리고 생균수의 경시적 변화는 최소증식저지농도에서 MIX,2MIC에서 24시간동안 계속적으로 감소되었다. 김치에 각 에스트리들을 0.1% 첨가함으로써 ascorbyl-6-laurate(??)가 가장 젖산생성을 억제하였다. 그리고 ??를 0.1%첨가한 경우 젖산 생성균도 현저한 저지효과를 나타내었다. Ascorbyl fatty acid esters(AFAE) were synthesized with ascorbic acid and caprylic, capric, lauric, myristic, and palmitic acid, respectively. The results confirmation tests of AFAE carried out, and antimicrobial activity of AFAE and possibities of using AFAE as food preservatives evaluated are summarized as follows: It was confirmed that purity of synthesized AFAE was satisfactory as demonstrated by GC/MS, IR< NMR< TLC and fatty acids reacted with -OH at No. 6 carbon of ascorbic acid to form ascorbyl fatty acid esters. As carbon chain of fatty acid was increased, melting point of AFAE became higher, whereas solubility lower. AFAE were decomposed by heating at 100℃, 30 min., indicating rather poor themostability. AFAE in general had stronger antimicrobial activity to gram positive than gram negative bacteria and ascorbyl-6-laurate demonstrated the strongest antimicrobial activity among the tested AFAE. Bacillus subtilis, Staphylococcus aureus and Lactobacillus brevis were most senitive to AFAE among the tested microorganisms, but Escherichia coli was not affected even at 500㎍/ml of AFAE. Numbers of cells were continuously decreased during incubation at MIC(minimal inhibitory concentration) or 2×MIC of AFAE. When 0.1% of each esters added to Kimchi, assorble-6-laurate could be inhibited effectively producing lactic acid and increase in cell number of lactic acid producing bateria was not remarkable.
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정동효,정호권,박준희,심상국 中央大學校 遺傳工學硏究所 1988 遺傳工學硏究論集 Vol.1 No.1
본 연구는 lac'z gene의 promoter개발을 위하여 착수하였다. lac'z gene의 promoterⅠ과 Ⅱ를 효모의 염색체 Bam HI DNA 단편에서 발견하였다. PromoterⅠ의 크기의 2.5kb정도이고 β- galactosidase 활성은 124.6 U/㎎ protein이었으나 promoterⅡ의 크기와 효소활성은 4.0kb 와 168.8 U/㎎이었다. 형질전환체에서 YEp plasmid 안정성은 52.7%에서 67.4%정도였다. YEp plasmid에서 YIp plasmid를 구출할 수 있었고 이 YIp plasmid는 대장균에서나 효모에서도 발현되었고 특히 HIS5 gene이 promoter로서 가능함을 알 수 있었다. It was attempted to observe the development of promoter on lac'z gene. Two promoterⅠand Ⅱ of lac'z gene were isolated from chromosomal DNA Bam HI fragment of yeast. The size of the promoterⅠwas estimated to be 2.5kb and β- galactosidase activity was 124.6 U/㎎ protein but the size of the promoterⅡ was 4.0kb 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. And YIp plasmid was constructed by YEp plasmid. This YIp plasmid both in E.coli and yeast. The HIS5 gene, coding gene of histidiolphosphate aminotransferase functioned as the promoter of YIp plasmid .
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