• Journal of Microbiology and Biotechnology
• /
• v.10 no.3
• /
• pp.287-292
• /
• 2000

Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.5
• /
• pp.509-516
• /
• 1998

A plasmid vector, pXGPRMATG-lac-Tgx, was developed for overproduction of $\beta$-galactosidase in Escherichia coli using the genetic components of groEx, a heat-shock gene cloned from symbiotic X-bacteria in Amoeba proteus. The vector is composed of intragenic promoters P3 and P4 of groEx, the structural gene of lac operon, transcription tenninator signals of lac and groEx, and ColEl and amp'of pBluescript SKII. The optimized host, E. coli DH5$\alpha$, transfonned with the vector constitutively produced 117,310-171,961 Miller units of $\beta$-galactosidase per mg protein in crude extract. The amount of enzyme in crude extract was 53% of total water-soluble proteins. About 43% of the enzyme could be purified to a specific activity of 322,249 Miller units/mg protein after two-fold purification, using two cycles of precipitation with ammonium sulfate and one step of gel filtration. Thus, the expression system developed in this study presents a low-cost and simple method for purifying overproduced $\beta$-galactosidase in E. coli.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.206-209
• /
• 2001

Recombinant human GM -CSF was expressed and secreted from transgenic rice cell suspension cultures in its biologically active form. This was accomplished by transforming rice callus tissues with an expression vector, pMYN44. containing the hGM -CSF cDNA. Regulated expression and secretion of hGM -CSF from this vector achieved using the promoter, signal peptide, and terminator from a rice alfa-amylase gene Amy3D. The Amy3D gene is expressed in response to sugar deprivation. The recombinant hGM -CSF was expressed from the transgenic rice cell culture on the sugar-free medium as a yield of about 110 mg/L in the culture filtrate, which was determined by ELISA. Biological activity of hGM-CSF was confirmed by measuring the proliferation of the hGM -CSF dependent TF -1 cells.(This work was supported by a grant from the NRL program of the Korean Ministry of Science and Technology. Shin, Y.- J.. Lee. J.-H and Kwon, T.-H. have been supported by BK21 program from the Korean Ministry of Education)

• Journal of The Korean Astronomical Society
• /
• v.34 no.1
• /
• pp.25-29
• /
• 2001

We have constructed a non-spherical model for the hot oxygen corona of Mars by including the effects of planetary rotation and diurnal variation of the Martian ionosphere. Exospheric oxygen densities are calculated by integrating ensemble of ballistic and escaping oxygen atoms from the exobase over the entire planet. The hot oxygen atoms are produced by dissociative recombination of $O^+_2$, the major ion in the Martian ionosphere. The densities of hot oxygen atoms at the exobase are estimated from electron densities which have been measured to vary with solar zenith angle. Our model shows that the density difference of hot oxygen atoms between noon and terminator is about two orders of magnitude near the exobase, but reduces abruptly around altitudes of 2000 km due to lateral transport. The diurnal variation of hot oxygen densities remains significant up to the altitude of 10000 km. The diurnal variation of the hot oxygen corona should thus be considered when the upcoming Nozomi measurements are analyzed. The non-spherical model of the hot oxy-gen corona may contribute to building sophisticate solar wind interaction models and thus result in more accurate escaping rate of oxygens from Mars.

• Journal of Microbiology and Biotechnology
• /
• v.5 no.3
• /
• pp.117-124
• /
• 1995

A gene (xynA) encoding the endo-xylanase (E.C.3.2.1.8) from Bacillus stearothermophilus was cloned in E. coli, and its complete nucleotide sequence was determined. The xynA gene consists of a 636 base pairs open reading frame coding for a protein of 212 amino acids with a deduced molecular weight of 23, 283 Da. A putative signal sequence of 27 amino acid residues shows the features comparable with the Bacillus signal sequences; namely, the signal contains a positively charged region close to the N-terminus followed by a long hydrophobic string. The coding sequence is preceded by a possible ribosome binding site with a free energy value of -16.6 kcal/mol and the transcription initiation signals are located further upstream. The translation termination codon (TAA) at the 3 end of the coding sequence is followed by two palindrome sequences, one of which is thought to act as a terminator. The xynA gene has a high GC content, especially in the wobble position of codons (64%). Comparison of the primary protein sequence with those of other xylanases shows a high homology to the xylanases belonging to family G.

• Journal of Sericultural and Entomological Science
• /
• v.39 no.2
• /
• pp.180-185
• /
• 1997

To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

• The Journal of Korean Institute of Communications and Information Sciences
• /
• v.24 no.6B
• /
• pp.1059-1067
• /
• 1999

In this paper characteristics of ATM-PON is described and a TC layer controller for ATM-PON OLT is designed. This paper proposes an algorithm of economically controlling the grant information written by CPU to dynamically allocate the upstream bandwidth on the PON among the ONUs in a fair way and of encrypting the downstream data using a lower standard of encryption, termed data churning, which is used to distinguish it from existing transmission system scramblers and higher layer encryption methods. This paper also proposes a method of allocating churning-related message into the PLOAM cell in order to synchronously change the churning key between the OLT and the ONU.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.2
• /
• pp.375-382
• /
• 2010

The available techniques for heterologous protein secretion in Lactobacillus strains are limited. The aim of the present study was to develop an efficient protein-secretion system using recombinant lactobacilli for various applications such as live delivery of biotherapeutics. For the construction of expression vectors, the Lactobacillus brevis slpA promoter, Lactobacillus casei prtP signal sequence, and mouse IL-10 sequences were used as a model system. Interestingly, the slpA promoter exhibited strong activity in L. casei, contrary to previous observations. In order to stabilize replication of the plasmid in E. coli, a removable terminator sequence was built into the promoter region. For the improvement of secretion efficiency, a DTNSD oligopeptide was added to the cleavage site of signal peptidase. The resulting plasmids provided remarkably efficient IL-10 secretion. Accumulation of the protein in the culture supernatant varied widely according to the pH conditions. By analysis of the secreted protein, formation of homodimers, and biological activity, IL-10 was confirmed to be functional. The presently constructed plasmids could be useful tools for heterologous protein secretion in L. casei.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
• /
• 2001.07a
• /
• pp.886-889
• /
• 2001

Silicone rubber is being used for the housing material of outdoor high voltage insulators such as composite insulator, bushing, surge arrestor and cable terminator because of good tracking and erosion resistance, good hydrophobicity and recovery of hydrophobicity and chemical stability. But, the leakge current occurs on surface of the composite polymeric insulation materials when the insulator is used for a long time with severe contaminative condition and it can lead the contamination flashover. So the leakage current is important to estimate the condition of the silicone rubber surface. In this paper, aging characteristics of silicone rubber used for outdoor insulation have been hydrophobicity of silicone rubber in salt fog chamber with average leakage current monitoring for observing the transformation of surface degradation properties of silicone rubber with different ATH(alumina trihydrate, A1$_2$O$_3$$.$3H$_2$O) filler contents. The experimental results show that a higher peak leakage current and to raise a long time for tracking with increasing amount of ATH by the salt fog and heat recycle ageing.

• Journal of Korea Society of Industrial Information Systems
• /
• v.17 no.3
• /
• pp.51-57
• /
• 2012

Pacman is known as the terminator of the loved classic game in the world. In this paper, classic pacman game without using a keyboard or a mouse we should be able to play the game with simple hand gestures. In other words, It use motion game to be a substitute for the hand direction key using the center coordinates. Also, movements of the monster is to be replaced depending on your hand movements by extracting a pointer of hand taking pictures in the MFC dialog using Cam. In this paper, YCbCbr image is convert to RGB images to extract skin color, and multiplication operations and hybrid median filtering was used in order to obtain better images. And it is used to obtain movement of the hand area based on obtain the center of gravity of the hand region.