• Korean Journal of Animal Reproduction
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• v.18 no.3
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• pp.191-197
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• 1994

It is generally known that mutations in any of several genes encoding photoreceptor-specific proteins have resulted in retinitis pigmentosa (RP), a disease characterized by losing photoreceptor function with progressive degeneration of photoreceptor cells and eventually leading to blindness. To study the procure and cure of photoreceptor degeneration, we produced transgenic mice. Transgene consisted of a 12.5kb genomic DNA fragment that contains mutated pig rhodopsin gene (Pro-347-Ser) including both the 5'-franking (4.0 kb) and the 3'-franking (2.9 kb) sequences. This gene was used for the production of transgenic mouse. The mutated rhodopsin DNA was microinjected into male pronuclei of fertilized mouse (C57BL /6]) embryos. We detected transgenic animals harboring mutated rhodopsin gene by PCR and Southern blot analysis. These transgenic mice showed stable transmission of microinjected rhodopsin gene into their offspring. Therefore these animals will provide a novel approach to study the mechanism of the photoreceptor degeneration and be provided as a disease model for the treatment of the blind in human.

• The Journal of Natural Sciences
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• v.5 no.1
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• pp.37-45
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• 1992

The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.1
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• pp.46-54
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• 1997

Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by malting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB (spo 5)1. We futher analyzed six recombinant plasmids, pDB (spo 5)2, pDB(spo 5)3, pDB(spo 5)4, pDB(spo 5)5, pDB(spo 5)6, pDB(spo 5)7, and found each plasmids is able to rescue the spo 5-2, spo 5-3, spo 5-4, spo 5-5, spo 5-6, spo 5-7, mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB (spo 5)1, and pDB (spo 5)R1 contained the spo 5 gene. Transcipts of spo 5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 25 kb were detected with 5 kb Hind III fragment containing a part of the spo 5 gene as a probe. The small mRNA (2.5 kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2 kb) was produced constitutively. Appearance of a 2.5 kb spo 5-mRNA depends upon the function of the mei1, mei2 and mei3 genes.

• Reproductive and Developmental Biology
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• v.41 no.1
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• pp.7-16
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• 2017

The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the ${\beta}-casein$ gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5' arm region and 1.8 kb or 0.64 kb of 3' arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5' terminal of endostatin gene and inserted into exon 7 of the ${\beta}-casein$ gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3' arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine ${\beta}-casein$ gene in the mammary gland.

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.16 no.6
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• pp.509-518
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• 2023

In this paper, we designed a 4Kb poly-fuse OTP IP (Intellectual Property) required for analog circuit trimming and calibration. In order to reduce the BL resistance of the poly-fuse OTP cell, which consists of an NMOS select transistor and a poly-fuse link, the BL stacked metal 2 and metal 3. In order to reduce BL routing resistance, the 4Kb cells are divided into two sub-block cell arrays of 64 rows × 32 rows, with the BL drive circuit located between the two 2Kb sub-block cell arrays, which are split into top and bottom. On the other hand, in this paper, we propose a core circuit for an OTP cell that uses one poly-fuse link to one select transistor. In addition, in the early stages of OTP IP development, we proposed a data sensing circuit that considers the case where the resistance of the unprogrammed poly-fuse can be up to 5kΩ. It also reduces the current flowing through an unprogrammed poly-fuse link in read mode to 138㎂ or less. The poly-fuse OTP cell size designed with DB HiTek 90nm CMOS process is 11.43㎛ × 2.88㎛ (=32.9184㎛2), and the 4Kb poly-fuse OTP IP size is 432.442㎛ × 524.6㎛ (=0.227mm²).

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.829-835
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• 2000

The ptsH and ptsI genes of Lactococus lactis subsp. lactis ATCC 7962 (L. lactis 7962), encoding the general proteins of phosphotransferase system (PTS) components, HPr and enzyme I, respectively, were cloned and characterized. A 1.3 kb PCR product was obtained using a primer set that was hybridized to the internal region of the L. lactis 7962 pts HI genes and then subcloned into a low-copy number vector, pACYC184. The 5' upstream and 3' downstream region from the 1.3 kb fragment were subsequently clone using the chromosome walking method. The complete ptsHI operon was constructed and the nucleotide sequences determined. Two ORFs corresponding to HPr (88 amino acids) and enzyme I (575 amino acids) were located. The ptsHI genes of L. lactis 7962 showed a very high homology (84-90%) with those genes from other Gram-positive bacteria. A primer extension analysis showed that the transcription started at either one of two adjacent bases upstream of the start codon. Using a Northern analysis, two transcripts were detected; the first, a 0.3 kb transcript corresponding to ptsH and the second, a 2 kb transcript corresponding to ptsH and ptsI. The transcription level of ptsH was higher than that of ptsI. The concentration of the ptsH transcript in cells grown on glucose was similar to that in cells grown on lactose, yet higher than that in cells grown on galactose. The ptsI transcript was scarcely detected in cell grown on lactose or galactose. The ptsI transcript was scarcely detected in cells grown on lactose or galactose. The results of a sequence analysis and Northern blot confirmed that the ptsH and ptsI genes of L. lactis 7962 were arranged in an operon like other known ptsHI genes and the expression of the ptsHI genes was regulated at the transcriptional level in response to the carbon source.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.205-209
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• 2008

This study was carried out to develop knock-in vector for EGFP (enhanced green fluorescent protein) expression in porcine $\beta$-casein locus. For construction of knock-in vector using porcine $\beta$-casein gene, we cloned the $\beta$-casein genome DNA from porcine fetal fibroblast cells, EGFP and SV40 polyA signal using PCR. The knock-in vectors consisted of a 5-kb fragment as the 5' recombination arm and a 2.7-kb fragment as the 3' recombination arm. We used the neomycin resistance gene ($neo^{r}$) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. To demonstrate EGFP expression from knock-in vector, we are transfected knock-in vector that has EGFP gene in murine mammary epithelial cell line HC11 cells with pSV2 neo plasmid. The EGFP expression was detected in HC11 cells transfected knock-in vector. This result demonstrates that this knock-in vector may be used for the development of knock-in transgenic pig.

• Korean Journal of Plant Resources
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• v.30 no.6
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• pp.679-685
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• 2017

Stachys affinis tubers are known for its high content of stachyose and eaten as an edible vegetable. In this work, we assessed on the anti-inflammatory and anti-proliferation activity of a various type of extracts derived from S. affinis tubers. The n-hexane and dichloromethane fractions were showed the high cytotoxicity on the cell lines including RAW264.7 macrophages, HEK293 human kidney cell, A549 human lung cancer cell, KB human oral cancer cell, and a PC-3 human prostate cancer cell. N-butanol and water fractions were not exhibited cytotoxicity on the tested cancer cells, limited in anti-inflammatory and anti-cancer activities. Nevertheless, the ethyl acetate fraction showed little harm to RAW264.7 cells but inhibited the production of nitric oxide (NO) and prostaglandin E2 (PGE2) significantly. In addition, it arrests the cell growth in A549, KB, and PC-3 cell while little cytotoxicity on HEK293 cells. Consequently, these results supported that the ethyl acetate fraction of S. affinis tubers could be a potential anti-inflammatory and anti-cancer ingredient.

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.10 no.6
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• pp.558-569
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• 2017

In this paper, a 256Kb EEPROM IP aimed at battery applications using a single supply of 1.5V which is embedded into an MCU is designed. In the conventional cross-coupled VPP (boosted voltage) charge pump using a body-potential biasing circuit, cross-coupled PMOS devices of 5V in it can be broken by the junction or gate oxide breakdown due to a high voltage of 8.53V applied to them in exiting the program or erase mode. Since each pumping node is precharged to the input voltage of the pumping stage at the same time that the output node is precharged to VDD in the cross-coupled charge pump, a high voltage of above 5.5V is prevented from being applied to them and thus the breakdown does not occur. Also, all erase, even program, odd program, and all program modes are supported to reduce the times of erasing and programming 256 kilo bits of cells. Furthermore, disturbance test time is also reduced since disturbance is applied to all the 256 kilo bits of EEPROM cells at once in the cell disturb test modes to reduce the cell disturbance testing time. Lastly, a CG driver with a short disable time to meet the cycle time of 40ns in the erase-verify-read mode is newly proposed.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.309-319
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• 2018

Previously, six Schizosaccharomyce pombe mutants that induce pheromone even in the presence of nitrogen source were isolated from a bank of temperature sensitive mutants. In this report, one of these mutants, pws6 was further characterized. The pheromone induction in pws6 mutant cells was specific to nutrient: the M-factor pheromone was induced without nitrogen starvation but not without glucose starvation. This result suggests that the pws6 mutant might have a specific defect in the pathway for nitrogen starvation. The pws6 mutant induces P-factor pheromone as well as M-factor without starvation of nitrogen in temperature sensitive mode, suggesting that the pheromone induction phenotype of pws6 mutation is not cell-type specific. From cloning of the $pws6^+$ gene by complementation of the temperature sensitive growth defect, three plasmids containing 8.1 kb, 3.3 kb, and 4.8 kb yeast DNA were recovered. These plasmids complement the growth defect of the pws6 mutant by 100%, 70%, and 10~20%, respectively. The abilities of these plasmids to complement pheromone induction phenotype of pws6 mutant cells were correlated well with the efficiencies of complementation of the growth defect. With comparison of their open reading frames to the complementation efficiencies, it is concluded that the open reading frame, SPBC19C7.06 is responsible for the complementation of temperature sensitive phenotype of the pws6 mutant. This open reading frame, named prs1, contains one long exon with no intron and encodes a putative prolyl tRNA synthetase. The putative Prs1 protein exhibits significant similarities to the prolyl tRNA synthetases of other species.