• Current Research on Agriculture and Life Sciences
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• v.4
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• pp.36-41
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• 1986

The transformation of Saccharomyces cerevisiae with YIp26, YRp7 and YEp13 was investigated. Transformation frequences of YIp5, YIp26, YRp7 and YEp13 in Escherichia coli HB101 was $5.1{\times}10^{-4}$, $1.5 {\times}10^{-3}$, $1.3{\times}10^{-3}$, $3{\times}10^{-3}$, respectively. When plasmids were used in covalently closed circular form, transformation frequency of YEp13 was $1.2{\times}10^{-4}$ in S. cerevisiae DBY747 and $3.3{\times}10^{-4}$ in S. cerevisiae MC16 and that of YRp7, YIp26 was $3{\times}10^{-6}$, below $6{\times}10^{-8}$ respectively in S. cerevisiae DBY747 by the method of Ito. Cotransformation of YIp26 and YEp13 in linear form increased the frequency of transformation with efficiences 270-fold higher than transformation of YIp26 only in S. cerevisiae DBY747. In cotransformation of YIp5+YEp13 and YIp26+YRp7 with S. cerevisiae DBY747 by Beggs' method. Expression frequency of YIp5+YEp13 and YIp26+YRp7 was $4{\times}10^{-6}$, $1.5{\times}10^{-6}$, respectively. The recombinant plasmid of cotransformant was thought that YIp26 and YEp13, YIp5 and YEp13, and YIp26 and YRp 7 were ligated in vivo in S. cerevisiae DBY747.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.48-53
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• 1993

For the purpose to improve the glucoamylase productivity of Saccharomyces diastaticus, we integrated STA 1 gene into chromosomal DNA of S. diastaticus using YIp vector. After construction of Ylp-STA by the subcloning of STAI (5.3 kb) into YIp5 vector, S. diastaticus GMT-II(a. ura3. STAJ) was transformed by Ylp-STA through homologous recombination at the chromosomal STAJ gene. So we obtained the tram formants that glucoamylase productivity was increased maximum six fold. These strains transformed by the multi-copy integration of Ylp-STA in chromosomal DNA were confirmed by Southern hybridization. And the integrated Ylp-STA was maintained stably during 30 mitotic divisions.

• Current Research on Agriculture and Life Sciences
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• v.5
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• pp.52-58
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• 1987

The yeast S. cerevisiae DBY747 was transformed with E. C - S. C shuttle vector YIp5, YEp13 and YRp7 by the method of spheroplast. The transformation frequency of YEp13 and YRp7 in S. cerevisiae DBY747 was $1.2{\times}10^3$ and $1.0{\times}10^2$ per $10{\mu}g$ of DNA, respectively. The transformants with YIp5 plasmid incapable of autonomous replication in S. cerevisiae were not detected in the condition of this experiment, but YIpS plasmid expressed the gene carried on it when cotransformed with a helper plasmid such as YEp13 or YRp7 : autonomously replicating plasmid. When plasmids were used in covalently closed circular form, cotransformation frequency of Ylp5-YEpl3 and Ylp5-YRp7 was 210 and 95 per $10{\mu}g$ of DNA, respectively. In cotransformation of linear plasmids, transformation frequency of the same cohesive ends was similar to that of noncomplementary cohesive ends. Transformants by the cotransformation with circular plasmids have been shown much higher frequency than with linear plasmids in S. cerevisiae DBY 747. The mitotic segregation stability test suggested that the cotransformant of YIpS-YEp13 was more stable than that of YIpS-YRp7.

• Journal of Life Science
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• v.28 no.11
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• pp.1277-1284
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• 2018

In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.

• Applied Biological Chemistry
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• v.38 no.1
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• pp.7-12
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• 1995

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 ${\beta}-galactosidase$ activity was 124.6 U/mg protein, and the size of the promoter II was 4.0 kb and its ${\beta}-galactosidase$ activity was 168.8 U/mg 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-lated from yeast chromosomal DNA can be used for promoter of plasmid YEp and YIp.

• The Journal of Korean Medical History
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• v.14 no.2
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• pp.189-248
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• 2001

In this essay, following final conclusions have been drawn by analyzing medical ideology and research system of Yi cheon in Yi hak yip mun("醫學入門"). Firstly, even though the existing medical history researchers are not noting the system of Yi-cheon's medical ideology, this essay has proved the man as a doctor who succeeded the (main system) based on the content of Yi-hak-yip-mun("醫學入門") by Ju-Dan-Gae(朱丹溪). The outline of this proof is as follows. 1. Those doctors who had actively researched in Myung era(明代), were basically taking over the medical studies and result of Gum-Won era(金元代). However, depending on whose theory is to be followed, the followers are largely divided into two groups of Ju-Dan-Gae Academics(丹溪學派) and On-Boe Academics(溫補學派). In addition, both Ju-Dan-Gae Academics(丹溪學派) and On-Boe Academics(溫補學派) hold contradictory ideologies to that of the main medical system. In Yi-hak-yip-mun("醫學入門"), Yi-cheon(李?) ties The Text of Whang-Jae-Nae-Kyung("黃帝內經"), Jang-Jung-Kyung(張仲景), Yu-Ha-Gun(劉河間), Yi-Dong-Won(李東垣), Ju-Dan-Gae(朱丹溪) into one pedigree. With regard to the main system, he especially marks Ju-Dan-Gae(朱丹溪) for his efforts in gathering various medical theories into a large compilement. 2. When Yi-Cheon(李?) was writing Yi-Hak-Yip-Mun("醫學入門"), he made references to various medical publishings, among those book which he had utilized, books by Ju-Dan-Gae Academics(丹溪學派) had affected him more than anything else in terms of both quality and quantity. 3. Yi-Cheon(李?)'s "Congested Phlegm Theory(痰鬱論)" had succeeded "Congested Phlegm Theory(痰鬱論) of Ju-Dan-Gae Academics(丹溪學派). His Yi-Hak-Yip-Mun("醫學入門"), carries a more complete form of "Congested Phlegm Theroy(痰鬱論) which was made into a more systemic and widely applicable method which was by Ju-Dan-Gae Academics(丹溪學派). Secondly, Yi-Hak-Yip-Mun("醫學入門"), is a medical book which was written in the process of systemic reorganization of medical theories of various academic parties in Myung 명 era. Since this process was hearing its completion in the period of Yi-Cheon(李?), he chose specific ways of reshuffling, whilst seeking ways to efficiently utilizing existing medical information . He provided a standard to specific ways. He rearranged the existing medical theories based upon these standards. He also contributed to clinical medicine by providing description of symptoms focused upon the symptoms differentiated In Conclusion, Yi-Hak-Yip-Mun("醫學入門") holds systematic medical information which was developed by Ju-Dan-Gae Academics(丹溪學派). Also, Yi-Cheon(李?) uniformly classified the clinical experiences of existing Ju-Dan-Gae Academics(丹溪學派). He had contributed in the clinical use of Ju-Dan-Gae Academic(丹溪學派)'s clinical experience by providing main points from differentiation of symptoms.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.200-204
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• 2013

To contruct amylolytic industrial strains of Saccharomyces cerevisiae which produce ethanol efficiently from raw starch, the Bacillus amyloliquefaciens ${\alpha}$-amylase genes (Amy) or Aspergillus awamori glucoamylase genes (GA1) was separately introduced into the ribosomal DNA loci in the chromosomes of the raw starch fermenting-parental strain (ATCC 9763/$YIp{\delta}AGSA{\delta}$), using double 18S rDNA-integration system. Ethanol production after 3 days of fermentation by the strain that produced ethanol most efficiently from raw starch (ATCC 9763/$YIp{\delta}AGSA{\delta}$/YIpAG2rD) among the transformant strains was 1.5-times higher than that by the parental strain. This new strain generated 9.2% (v/v) ethanol (72 g/L) from 20% (w/v) raw corn starch and consumed 75% of the raw starch content during the same period.

• The Korea Genome Conference
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• 2003.09a
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• pp.60.1-60.1
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• 2003

• Journal of Digestive Cancer Research
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• v.5 no.1
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• pp.1-4
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• 2017

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.215-221
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• 1991

An autonomous replication sequence (ARS) derived from Cephalosporium acremonium ATCC 20339 was cloned in Sarchuromyces cerevisiae SHY 3 using YIp5 as a cloning vector. A new recombinant plasmid, designated pCY-2, which contained a 3.7 kb BamHI fragment of C. acrenzonium DNA showed the highest stability among the 40 recombinant plasmids composed of the YIp5 2nd ARS of C. ucremoniztm. Also, Southern hybridization and transformation of E, cull with DNA purified from yeast transformants verified that pCY-2 autonomously replicates in yeasts. Transformation efficiency and plasmid stability of pCY-2 in yeast were higher than those ol YRp 7 containing ARS which originated from yeast. Detailed studies by subcloning revealed that two ARSs existed within 2.6 kb of the insert, which is a novel discovery. However, it was concluded that these two ARSs were ligated during the gene manipulation in vitro.