• Journal of Plant Biotechnology
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• 제4권2호
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• pp.59-65
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• 2002

ADP-glucose pyrophosphorylase (AGPase) catalyzes the key regulatory step in starch biosynthesis. Two cDNA clones encoding AGPase subunits were isolated from the leaf cDNA library of Chinese cabbage (Brassica campestris L. spp. pekinensis). One was designated as BCAGPS for the small subunit and the other as BCAGPL for the large subunit. Both cDNAs have uninterrupted open reading frames deriving 57 kDa and 63 kDa polypeptides for BCAGPS and BCAGPL, respectively, which showed significant similarity to those of other dicot plants. Also, However, the deduced amino acid sequence of BCAGPL has a unique feature. That is, it contains two regions (Rl and R2) lacking in all other plant enzymes. This is the first report of BCAGPL containing Rl and R2 among plant large subunits as well as small subunits. From the genomic Southern analysis and BAC library screening, we inferred the genomic status of BCAGPS and BCAGPL gene.

• Plant Biotechnology Reports
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• 제2권4호
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• pp.253-258
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• 2008

The cDNA of the touch-induced genes (TCH) of the sweet potato [Ipomoea batatas (L.) Lam.] has been cloned and analyzed. IbTCH1, which exists as at least two-copy genes in the genome of the sweet potato, encodes for 148-amino acid polypeptides, and harbors four conversed $Ca^{2+}-binding$ motif EF-hands. IbTCH1 was shown to be expressed in the flower, leaf, thick pigmented root, and particularly in the white fibrous root, but expressed only weakly in the petiole. IbTCH1 is upregulated upon exposure to environmental stresses, dehydration, and jasmonic acid. Furthermore, IbTCH1 is developmentally regulated in the leaf and root. These results strongly indicate that the gene performs functions in both plant development and in defense/stress-signaling pathways.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.579-586
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• 2005

When subjected to hyperosmotic pressure by NaCl addition, H69K-NGD transfectoma, like KR12H-2 transfectoma, displayed decreased specific growth rate (${\mu}$) and increased specific antibody productivity ($q_{Ab}$): Elevation of medium osmolality from 280 mOsm/kg to 415 mOsm/kg decreased ${\mu}$ by $79\%$ in batch cultures of H69K-NGD transfectoma, while it increased $q_{Ab}$ by $103\%$. However, unlike KR12H-2 tranfectoma, enhanced $q_{Ab}$ of H69K-NGD transfectoma at hyperosmolalities was not due to elevated levels of Ig mRNAs. In hyperosmotic cultures of H69K-NGD transfectoma, heavy-chain mRNA per cell was not enhanced with increasing osmolality. Hyperosmotic pressure was found to preferentially enhance immunoglobulin (Ig) translation rates of H69K-NGD transfectoma. However, under hyperosmotic pressure, the translation rate of Ig polypeptides was not enhanced as much as $q_{Ab}$. This result suggests that hyperosmotic pressure also influences the post-translational process. Taken together, the results obtained show that intracellular response of transfectomas to hyperosmotic pressure, in regard to the main intracellular steps of the antibody secretory pathway, is cell-line dependent.

• Fisheries and Aquatic Sciences
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• 제20권9호
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• pp.23.1-23.9
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• 2017

Two distinct cDNAs encoding aquaporins (mmAQPs 1a and 3a) were isolated and characterized from mud loach Misgurnus mizolepis. The identified mud loach AQP cDNAs encode for polypeptides of 260 and 302 amino acids. Topology predictions confirmed six putative membrane-spanning domains connected by five loops and the N- and C-terminal domains being cytoplasmic. The mud loach AQPs 1a and 3a showed broad distribution in multiple tissues including immune-responsive tissues as well as osmoregulatory tissues. Hence, the diversity of AQP distribution and expression possibly indicated its differential functions in the regulation of fluid movement in response to environmental stimuli. The transcription of mmAQP genes was differentially modulated by immune challenges. In particular, the mmAQP3a expression level in the liver was more responsive to immune challenges than that of mmAQP1a. Taken together, fish stimulation or infection resulted in significant modulation of mud loach AQP genes, suggesting potential functional roles of these proteins in piscine pathophysiological process.

• 한국동물학회지
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• 제33권3호
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• pp.303-309
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• 1990

대장균에서 진핵세포의 펩타이드 호르몬 전구체를 대량으로 생산할 목적으로 T7 overexpression system을 이용하여 angler 어류의 prepro-SRIF I 유전자와 T7 phage coat 단백질 S10 유전자를 결합시켜 일련의 융합유전자를 합성하였다. 이 융합유전자를 가지고 있는 숙주 대장균, BL21 DE3는 3가지 종류의 서로 다른 SRIF 관련 폴리펩타이드를 대량 합성하였다. 본 연구에서는 대량합성된 폴리펩타이드의 특성을 규명하였으며, 펩타이드 호르몬 전구체를 얻기 어려운 이종이에서의 대량발현의 중요성과 표적 펩타이드 호르몬인 SRIF의 적용성을 논의하였다.

• 약학회지
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• 제47권3호
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• pp.184-189
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• 2003

Peptide deformylase (PDF) is essential and unique to bacteria, thus making it an attractive target for the discovery of novel antibacterial drugs. PDF deformylates the N-formylmethionine of newly synthesized polypeptides in prokaryotes. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and PDF protein was over-produced in Escherichia coli BL21 (DE3). NH$_2$-terminal His-tagged PDF protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. Enzymatic activity of purified 6xHis-tagged PDF was tested on the substrate (formyl-Methionine-Alanine-Serine) by formate dehydrogenase-coupled spectrometric assay of peptide deformylase. For the discovery of new PDF inhibitors from chemical libraries and culture broths of soil bacteria, a target-oriented screening system using a 96-well plate was developed. About 3,000 commercial chemical libraries were tested in this screening system, and 2 chemicals (0.07％) among them showed an inhibitory activity against PDF enzyme. This result showed that a new screening system can be used for the discovery of new PDF inhibitors.

• 대한약리학회지
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• 제11권1호
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• pp.15-18
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• 1975

The effect of skin extracts of Korean amphibian, poisonous snake and fresh-water fish were determined for their caerulein-like action on rabbit gall bladder strips. The isolated gall bladder strips were prepared according to the technique described by Amer and Becvar(1969). The strips were placed in a bath containing 100ml of Locke-Ringer solution maintained at $38^{\circ}C$. Oxygen was continuously bubbled through the solution. The tension of the muscle strip was initially adjusted to 0.7g. The contractile response was measured isometrically by a force-displacement transducer connected to a polygraph. In this rabbit gall bladder strip caerulein produced contraction of CCK-PZ type. The skin extract of Korean amphibian also elicited similar contraction as caerulein, which extracted from Australian amphibian, Hyla caerulea, by Erspamer et al. The calculated amount was approximately $2{\mu}g$ caerulein per gram of skin tissue in Korean amphibian and the potency was about 1/200 of that seen in Australian amphibian. The contraction of gall bladder strip by our amphibians occurs in decreasing order; Rana Nigromaculata coreana Okada, Rana nigromaculata Hallowell, Hyla arborea japonica Gunther and Bombina orientalis Boulenger. The skin extracts of poisonous snake and fresh-water fish produced no caerulein-like activity.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1983-1992
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• 2016

Pseudomonas strains isolated from the heavily contaminated Lubin copper mine and Zelazny Most post-flotation waste reservoir in Poland were screened for the presence of integrons. This analysis revealed that two strains carried homologous DNA regions composed of a gene encoding a DNA_BRE_C domain-containing tyrosine recombinase (with no significant sequence similarity to other integrases of integrons) plus a three-component array of putative integron gene cassettes. The predicted gene cassettes encode three putative polypeptides with homology to (i) transmembrane proteins, (ii) GCN5 family acetyltransferases, and (iii) hypothetical proteins of unknown function (homologous proteins are encoded by the gene cassettes of several class 1 integrons). Comparative sequence analyses identified three structural variants of these novel integron-like elements within the sequenced bacterial genomes. Analysis of their distribution revealed that they are found exclusively in strains of the genus Pseudomonas.

• 한국식품영양과학회지
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• 제24권3호
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• pp.362-370
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• 1995

Human plasma low density lipoprotein(LDL) is the main carrier for cholesterol, and recent studies suggest the normal LDL can be readily oxidized by free radical and not interact with LDL receptor. Lipoprotein pariticles are consisted of lipid andprotein, and fatty acids of lipoproteins are prone to oxidation. LDL particles readily undergo oxidative modification by copper. From the results, oxidized LDL altered its biological properties. A marked increase in the electrophoretic mobility of LDl on agarose gel indicated that negative surface charge of the LDL particles was increased. Also, the results from the HPLC showed that oxidized LDL was degraded into several polypeptides nonenzymatically. Degradation tests which measured the amount of 5-IAF labelled oxidized LDL were carried out by monocyte and hepatocyte cell culture. Hepatocyte cell culture of modified LDL did not show consistent pattern. However, binding rate of modified LDL with HMDM(human monocyte derived macrophage) was enhanced with oxidation, but was retarded by addition of antioxidants(hyaluronic acid, vitamin A, vitamin E). Also comparisons of oxidized-LDL, acetyl-LDL and MDA-LDL showed significant differences in the chemical properteis and binding affinity to HMDM. Thus, modificaition of normal LDL altered its biological properties.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권3호
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• pp.229-235
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• 2004

The influence of sucrose on in vitro growth, chlorophyll content, and rubisco/rubisco activase were studied in tobacco leaves. The most pronounced effect on in vitro growth and the chlorophyll content was found at 4% sucrose. The rubisco content increased with increasing concentrations of sucrose, but a point was reached beyond which the increasing concentrations of sucrose caused an inhibition of this enzyme. The rubisco activity showed patterns of change similar to the rubisco content. These data suggest that sucrose may have an affect on the activation and induction of rubisco and that sucrose can be both a positive effector and negative effector depend on its concentration. The degree of intensity of 55 and 15 kD polypeptides, which were identified as the large and small subunit of rubisco, respectively, by SDS-PAGE analysis at 4% sucrose was significantly higher than that of other treatments, indicating that sucrose had an effect on both subunits. We subsequently examined whether the rubisco content and activity of being induced by sucrose is associated with rubisco activase. The rubisco activase content at 4% sucrose was higher than that of other treatments. A similar change pattern was also observed in the activity of rubisco activase. The intensity of two 52 and 51 kD polypeptide bands at 4% sucrose was higher than that of corresponding bands of other treatments. The stimulatory and inhibitory effects of rubisco by sucrose seemed to be caused by rubisco activase.