• Journal of Photoscience
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• v.11 no.1
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• pp.41-45
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• 2004

The key riboflavin synthesis genes are located immediately downstream of luxG in the lux operon from Photobacterium leiognathi. It is of interest that a site capable of forming a rho-independent terminator does not appear to be present between luxG and ribE in our previous data. These results raise the question of whether the transcription of lux and rib genes is integrated or not. In order to answer the question, in vivo transcriptional assay and Southern blot were examined. These studies demonstrate that neither transcriptional terminator nor promoter site is present in the intergenic region between of lux and rib genes as well as that the riboflavin genes are single copy in a chromosome of Photobacterium leiognathi.

• Mycobiology
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• v.31 no.1
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• pp.42-45
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• 2003

For transformation of Pleurotus ostreatus, two novel vectors, pPhKM1 and pPhKM2, were constructed, using the regulatory sequences of the P. sajor-caju $\beta$-tubulin gene(TUB1) and the ble gene encoding phleomycin binding protein. pPhKM1 contains ble fused to the TUB1 promoter and the Schizophyllum commune GPD terminator. pPhKM2 contains ble fused to the promoter and terminator regions of P. sajor-caju TUB1. To confirm phleomycin-resistance activity, each vector was cotrans-formed with pTRura3-2 into the P. ostreatus homokaryotic $ura^-$ strain. The transforming DNA was stably integrated into the genomic DNA. Subsequently, phleomycin resistance was conferred on wild-type dikaryotic P. ostreatus by transformation with pPhKM1 or pPhKM2. This transformation system generated stable phleomycin-resistant transformants.

• BMB Reports
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• v.38 no.6
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• pp.690-694
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• 2005

Transcription termination of the human mitochondrial genome requires specific binding to termination factor mTERF. In this study, mTERF was produced in E. coli and purified by two-step chromatography. mTERF-binding DNA sequences were isolated from a pool of randomized sequences by the repeated selection of bound sequences by gel-mobility shift assay and polymerase chain reaction. Sequencing and comparison of the 23 isolated clones revealed a 16-bp consensus sequence of 5'-GTG$\b{TGGC}$AGANCCNGG-3' in the light-strand (underlined residues were absolutely conserved), which nicely matched the genomic 13-bp terminator sequence 5'-$\b{TGGC}$AGAGCCCGG-3'. Moreover, mTERF binding assays of heteroduplex and single-stranded DNAs showed mTERF recognized the light strand in preference to the heavy strand. The preferential binding of mTERF with the light-strand may explain its distinct orientation-dependent termination activity.

• Proceedings of the Optical Society of Korea Conference
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• 2005.02a
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• pp.192-193
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• 2005

• Applied Biological Chemistry
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• v.48 no.4
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• pp.335-338
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• 2005

For the development of qualitative PCR detection method of genetically modified rice (Oryza sativa L.), rice species specific gene, SAMDC1 (S-adenosylmethionine decarboxylase), was selected and validated as suitable for use as an endogenous reference gene in rice. The primer pair OsSAMDC1-5'/3' with 110 bp amplicon was used for amplification of the rice endogenous gene, SAMDC1 and no amplified product was observed from 19 different plants as templates. Qualitative PCR method was assayed with 2 different GM rices (Milyang 204 and Iksan 483) developed in Korea. For the qualitative PCRs, the construct-specific detection primer pairs were constructed. Os204-5'/OsNOS-3' amplifying the junction region of GUS gene and NOS terminator introduced in Milyang 204 gave rise to an amplicon 172 bp; also, Os483-5'/OsNOS-3' amplifying the junction region of Bar gene and NOS terminator introduced in Iksan 483 gave rise to an amplicon 161 bp.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.348-355
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• 2012

In the present study, we investigated the overexpression and characterization of bovine pancreatic (bp)- DNase I in Saccharomyces cerevisiae and Pichia pastoris. The bp-DNase I gene was fused in frame with the GAL10 promoter, $MF{\alpha}$, and GAL7 terminator sequences, resulting in the plasmid, pGAL-$MF{\alpha}$-DNaseI (6.4 kb). Also, the bp-DNase I gene was fused in frame with the AOX1 promoter, $MF{\alpha}$, and AOX1 terminator sequences, resulting in the plasmid, pPEXI (8.8 kb). The recombinant plasmids, pGAL-$MF{\alpha}$-DNaseI and pPEXI were introduced into S. cerevisiae and P. pastoris host cells, respectively. When the transformed yeast cells were cultured at $30^{\circ}C$ for 48 h in galactose or methanol medium, bp-DNase I was overexpressed and the most of activity was found in the extracellular fraction. P. pastoris transformant activity showed 45.5 unit/mL in the culture medium at 48 h cultivation, whereas S. cerevisiae transformant revealed 37.7 unit/mL in the extracellular fraction at 48 h cultivation. The enzymatic characteristics, such as DNA cleavage and half life were investigated. Treatment of the recombinant DNase I from P. pastoris induced degradation of the calf thymus DNA within 1 minute, and this DNA degradation rate was higher than that of commercial bp-DNase I (SIGMA) and the recombinant DNase I from S. cerevisiae.

• Bulletin of the Korean Chemical Society
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• v.32 no.6
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• pp.2129-2131
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• 2011

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.290.2-290
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• 1999

No Abstract, See Full Text

• Microbiology and Biotechnology Letters
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• v.25 no.6
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• pp.571-579
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• 1997

For the overproduction of pectate lyase (PL), the recombinant plasmid pl2BS fl which has strong promoter from alkali-tolerent Bacillus sp. YA-14 was used. In order to overexpress the pectate lyase by the action of overlapping strong promoter in pl2BS$\Delta$fl, 1.6 kb of PL gene was inserted into pl2BS$\Delta$fl to form pl2BS$\delta$f1-PL and the enzyme was expressed. But decreased expression efficiency of the PL gene was observed and it was due to the presence of the transcription terminator region on the upstream of the PL gene. The transcription terminator of the PL gene in pl2BS$\delta$f1-PL was removed and the resulting plasmid p12BS$\Delta$fl$\Delta$PL was formed. Bacillus subtilis 207-25 harboring the recombinant plasmid, p12BS$\Delta$fl$\Delta$PL, revealed increased expression efficiency with chloramphenicol induction when cat-86 was used as a reporter gene.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.348-355
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• 1991

DNA segment encoding $\beta$-1, 4-glucanase of Bacillus subtilis was fused in frame to mouse $\alpha$-amylase signal sequence behind the alcohol dehydrogenase isoenzyme I gene (ADHI) promoter of the yeast expression vector pMS12. To enhance the expression level of the $\beta$glucanase gene in yeast, transcription terminator sequence iso-1-cytochrome c gene (CYCI) was inserted into the recombinant plasmid. The transformants harbouring such recombinant plasmids secreted $\beta$-glucanase into the culture medium. The expresstion level of the $\beta$-glucanase gene was increased about 2-fold caused by inserting the terminator. The amount of the secreted $\beta$-glucanase in culture medium was approximately 60% of the total quantity synthesized.