• 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.

• The Korean Journal of Zoology
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• v.39 no.1
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• pp.115-121
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• 1996

To investigate gestational profiles in gene expression of placental lactogen I fpL4), PL-lI and Pit-1, RNA samples were extracted from the placentas of pregnant day 12 to 20 at 2 day intervals. Northern blots showed changes in gene expression of PL4, - 11 and Pit-i. Sizes of PL-l and -II mRNA were changed and amounts of PL-I, -H and Pit-1 mRNA increased during progress of gestation. To examine the effect of estrogen on the gene expression of PL-I, -Il and Pit-1, pregnant female rats were ovariectomized (OVX) and daily injected with estradiol (OVX + E). OVX markedly lowered the amount of PL4 and 41 mRNA, and shifted niRNA size from 1 kb to i 3 kb in PL-l mRNA and 0.6 kb to i kb in PL-ll mRNA, respectively. OVX had no effect on the mRNA size of Pit-1, but markedly attenuated Pit-1 mRNA level. Estrogen injection reversed the effect of OVX on the size-shift but not on the amount of PL4 and -Il mRNA. Replacement of E partially recovered OVX-induced inhibition of Pit-i mRNA level. Present results suggest that estrogen may play a pivotal role on the gene expression of PL-l and -Il such as alternative RNA splicing and/or polyadenylation, and Pit-1 may be involved in the gene expression of PL-l and 41 by estrogen.

• Korean Journal of Breeding Science
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• v.42 no.1
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• pp.50-55
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• 2010

The R-r:standard (R-r:std) allele of maize R gene complex consists of S subcomplex and P component; the S subcomplex regulates anthocyanin pigmentation of seed aleurone layer, and the P component confers pigmentation of the other plant parts. The S subcomplex contains two functional genes, S1 and S2 components. In the presence of Pl gene some alleles of R gene induce anthocyanin pigmentation of pericarp. In the present study, the effects of different R alleles on the anthocyanin pigmentation of pericarp in the presence of Pl gene were analyzed in order to identify the R gene component responsible for pericarp pigmentation. The results show that R-ch and r-ch alleles condition similar degrees of pericarp pigmentation, and that R-r:Ecuador (R-r:Ec) conditions stronger pigmentation. The r-ch allele, which is inferred that its S subcomplex has lost function but the P component is normal, induces pericarp pigmentation in the presence of Pl gene. On the contrary, the R-g:g1111 allele, derived from R-r:Ec and inferred that its S subcomplex functions normal but the P component has lost its function, did not induce pericarp pigmentation in the presence of Pl gene. Moreover, PCR analysis of genomic DNA's of R-ch and r-ch indicate that R-ch maintains both P and S1 components, whereas r-ch lacks for the S1 component. Taken together, The results suggest that the P components of R alleles inducing pericarp pigmentation in the presence of Pl gene are responsible for pericarp pigmentation.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1819-1826
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• 2015

Poly(ε-ʟ-lysine) (ε-PL) is a novel bioactive polymer secreted by filamentous bacteria. Owing to lack of a genetic system for most ε-PL-producing strains, very little research on enhancing ε-PL biosynthesis by genetic manipulation has been reported. In this study, an effective genetic system was established via intergeneric conjugal transfer for Streptomyces albulus PD-1, a famous ε-PL-producing strain. Using the established genetic system, the Vitreoscilla hemoglobin (VHb) gene was integrated into the chromosome of S. albulus PD-1 to alleviate oxygen limitation and to enhance the biosynthesis of ε-PL in submerged fermentation. Ultimately, the production of ε-PL increased from 22.7 g/l to 34.2 g/l after fed-batch culture in a 5 L bioreactor. Determination of the oxygen uptake rate, transcriptional level of ε-PL synthetase gene, and ATP level unveiled that the expression of VHb in S. albulus PD-1 enhanced ε-PL biosynthesis by improving respiration and ATP supply. To the best of our knowledge, this is the first report on enhancing ε-PL production by chromosomal integration of the VHb gene in an ε-PL-producing strain, and it will open a new avenue for ε-PL production.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.31 no.1
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• pp.27-34
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• 2009

Angiogenesis is essential for solid tumor growth and progression. Among the pro-angiogenetic factors, vascular endothelial growth factor(VEGF), also known as vascular permeability factor, is the most important as a mitogen for vascular endothelium. The VEGF family of molecules currently consists of six growth factors, VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, and placenta growth factor(PlGF). Over-expression of PlGF is associated with angiogenesis under pathological conditions such as ischemia, inflammation, and cancer. Hence, the goal of this study is to identify the correlation of clinicopathlogical factors and the up-regulation of PlGF expression in oral squamous cell carcinoma. We studied the immunohistochemical staining of PlGF, PlGF gene expression and a real time quantitative RT-PCR in 20 specimens of 20 patients with oral squamous cell carcinoma. The results were as follows. 1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, the high level staining of PlGF was observed. And the correlation between immunohistopathological PlGF expression and histological differentiation of specimens was significant (Pearson correlation analysis, significance [r] >0.6, P < .05). 2. In the PlGF gene RT-PCR analysis, PlGF expression was more in tumor tissue than in adjacent normal tissue. Paired-samples analysis determined the difference of PlGF mRNA expression level between the cancer tissue and the normal tissue (Student's t - test, P < .05) These findings suggest that up-regulation of the PlGF gene may play a role in progression and local metastasis in invasive oral squamous cell carcinoma.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.316-319
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• 1988

Pectate Lyase (PL) was cloned from alkali-tolerant Bacillus sp. YA-14 into Escherichia coli MB1000 by inserting HindIII-generated DNA fragment into the HindIII site of pBR322 and then screening recombinant transformant for the ability to hydrolyze sodium polypectate on agar plate, The recombinant plasmid, called pYPC29, was isolated, and the size of the cloned HindIII fragment was found to be 1.6 kb. The PL gene was stablely maintained and expressed efficiently in Escherichia coli. The Pt accumulated largely in the periplasmic space of Escherichia coli clones.

• KSBB Journal
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• v.13 no.4
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• pp.383-390
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• 1998

Bacillus subtilis DT134 was resistant to 50-fold higher concentration of cadmium ions (Cd2+) than cadmium-sensitive B. subtilis BD224 in Luria Broth (LB) medium. Minimal inhibition concentration test in LB agar plates also showed similar results. The elevated cadmium resistance of B. subtilis DT134 strongly suggested a possible existence of cadmium resistance gene in it. Southern blot with Staphylococcus aureus cadA gene fragment (757 bp NlaIV-XmnI cadA DNA fragment) as probe was carried out to test the existence and similarity of the gene. In high stringency condition, there was no detectable signal, but in low stringency, a strong signal specific to the cadA probe could be detected. These results strongly suggested that there was some similarity between total DNA of B. subtilis DT134 and S. aureus pl258 in terms of cadmium resistance gene and the resistance mechanism might be an efflux mechanism. The subsequent efflux experiment showed that the cadmium resistance mechanism of B. subtilis DT134 was also due to the efflux of cadmium.

• Applied Biological Chemistry
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• v.31 no.3
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• pp.219-225
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• 1988

Reverse transcriptase gene of Avian sarcoma virus(ASV) was cloned with a thermoinducible expression vector, pPL-lambda. E. coli N4830 which carries temperature sensitive cI857 la mbda repressor, was transformed with this pPL-pol plasmid DNA. The RNA transcribed by those tranoformants was isolated and analyzed. It was shown that the inserted reverse transcriptase gene of ASV was transcribed at high-level when cells were grown at high temperature.

• The Journal of Korean Obstetrics and Gynecology
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• v.22 no.2
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• pp.43-59
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• 2009

Purpose: This study was performed to determine the inhibitory effect of Persimmon Leaves extract (PL) on melanin synthesis in B16F10 melanoma cells B16F10. Methods: The inhibitory effects of PL on melanin synthesis were determined by in vitro assay. To elucidate inhibitory effects of PL on melanin synthesis, we determined the melanin release and melanin production in B16F10. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, PKA, PKC${\beta}$, ERK-1, ERK-2, AKT-1, MITF in B16F10. Results: 1. PL inhibited melanin release, melanin production in B16F10. 2. PL inhibited tyrosinase activity in vitro and in B16F10. 3. PL suppressed the expression of tyrosinase, TRP-1, TRP-2 in B16F10. 4. PL suppressed the expression of PKA, PKC${\beta}$ in B16F10. 5. PL increased the expression of ERK-1, ERK-2, AKT-1 in B16F10. 6. PL suppressed the expression of MITF in B16F10. Conclusion: From these results, it may be concluded that PL is possesed of the antimelanogenetic effects.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.91-91
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• 2003

We have studied the mechanism of action of TCDD on CYP1A1 promoter activity in both Hepa I and MCF-7 cells using transient transfection system with plAl-Luc reporter gene. When HDAC inhibitors, such as trichostatin A, HC toxin and a novel HDAC inhibitor, IN2001 were cotreated with TCDD to the cells transfected with plAl-Luc reporter gene, the basal promoter activity of CYP1A1 was increased by HDAC inhibitors. Also, in MCF-7 human breast cancer cells, HDAC inhibitors, such as IN2001 and trichostatin A increased the basal activity of CYP1A1 promoter but TCDD stimulated CYP1A1 promoter activity was not changed by HDAC inhibitors. And, in stably-transfected Hepa I cells with plAl-Luc, HDAC inhibitors increased the basal promoter activity only.