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Plasmid DNA pBR 322의 증폭과 Hydroxyapatite Chromatography에 의한 분리
오두환,유주현,신원철,정건섭,유승구 연세대학교 산업기술연구소 1984 논문집 Vol.16 No.1
The amplification and the isolation conditions of pBR322 plasmid DNA were investigated. For the amplification of pBR322 plasmid DNA, cells from logarithmic growth phase were most effective. The optimal conditions for the amplification of pBR322 plasmid DNA in Escherichia coli GM4 were obtained as follows; 50-150 ㎍/ml of chloramphenicol concentration, 6-8 hours of incubation time in the presence of chloramphenicol. Ampicillin and tetracycline had no effect on the amplification of pBR322 plasmid DNA. Approximately 1 mg of pBR322 plasmid DNA was purified from 3.5 ℓof Escherichia coli GM4 culture broth by hydroxyapatite chromatography. The purified pBR322 plasmid DNA was expressed in Escherichia coli C600.
윤기도,권동진,홍석산,김수일,정건섭 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4
To investigate the inhibitory effect of soybean and Korean traditional fermented soybean products on the chemically induced mutagenesis, we extracted soybean, Kanjang, Doenjang, Kochujang, and Chonkukjang with water, methanol and hexane. Inhibitory effect of extracts was assayed by the SOS chromotest using Escherichia coli PQ37 as a test strain. 4-nitroquinoline-1-oxide(4NQO), N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), and aflatoxin B_1(AFB_1) were used as mutagens. Methanol extracts showed relatively higher inhibitory effect than water and hexane extracts. Methanol extracts of soybea, Doenjang, Kochujang, and Chongkukjang showed inhibitory effect of 68.4, 96.3, 17.5, and 100.9%, against MNNG, and 28.6, 109.1, 41.3, and 101.8% against AFB_1., respectively. Doenjang methanol extract showed inhibitory effect of 51.0, 96.3, and 109.1% against 4NQO, MNNG, and AFB_1. Inhibitory effect of heat-treated Doenjang and Chongkukjang methanol extracts on the mutagenicity of MNNG and AFB_1 was remained over 95% of the inhibitory effect of heat-untreated extracts, demonstrating the heat stability of the potent antimutagenic activity.
Secretion of Bovine β-Casein by Saccharomyces cerevisiae
Chung, Kun Sub,Rafael, F. R.,Oh, Sang Suk,Richardson, T. 한국미생물 · 생명공학회 1991 Journal of microbiology and biotechnology Vol.1 No.1
Yeast expression plasmids containing an appropriate leader sequence and bovine β-casein cDNA were constructed to produce β-casein for the study of its functional characteristics. Two kinds of expression systems for β-casein were constructed using pCGY1444 as a precursor plasmid. This plasmid is a yeast-E. coli shuttle vector which contains the chelatin promoter. The plasmid pISB202 contains the invertase leader sequence and β-casein gene. The plasmid pDEB303 contains the original bovine β-casein leader sequence gene. These two plasmids were introduced into S. cerevisiae AB116 which is a strain deficient in the major yeast proteinases. Each clone was grown in minimal media for 24h before induction by CuSO_4. The cells were thus grown under expression conditions. Both strains harbouring pISB202 and pDEB303 expressed bovine β-casein. The β-casein was detected using immunochemical staining after western blot. Secretion of β-casein was detected in the culture broth. The estimated amount of secreted β-casein was approximately 50㎍/ℓ.
Secretion of Bovine $\beta$-Casein by Saccharomyces cerevisiae
Chung, Kun-Sub,Rafael, F.R.,Oh, Sang-Suk,Richardson, T. The Korean Society for Microbiology and Biotechnol 1991 Journal of microbiology and biotechnology Vol.1 No.1
Yeast expression plasmids containing an appropriate leader sequence and bovine $\beta$-casein cDNA were constructed to produce $\beta$-casein for the study of its functional characteristics. Two kinds of expression systems for $\beta$-casein were constructed using pCGYl444 as a precursor plasmid. This plasmid is a yeast-E. coli shuttle vector which contains the chelatin promoter. The plasmid pISB202 contains the invertase leader sequence and $\beta$-casein gene. The plasmid pDEB303 contains the original bovine $\beta$-casein leader sequence gene. These two plasmids were introduced into S. cerevisiae AB116 which is a strain deficient in the major yeast proteinases. Each clone was grown in minimal media for 24 h before induction by $CuSO_4$. The cells were thus grown under expression conditions. Both strains harbouring pISB202 and pDEB303 expressed bovine $\beta$-casein. The $\beta$-casein was detected using immunochemical staining after western blot. Secretion of $\beta$-casein was detected in the culture broth. The estimated amount of secreted $\beta$-casein was approximately 50 ${\MU}g$/l.