• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.26-31
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• 2023

A map gene encoding methionyl-specific aminopeptidase (MAP; EC 3.4.11.18) was cloned from Tetragenococcus halophilus CY54. Translated amino acid sequence of CY54 MAP showed high similarities with those from Enterococcus faecalis (83.8%) and Streptococcus salivarius (62.2%) but low similarities with MAPs from Lactobacillus and Lactococcus genera. The map gene was overexpressed in E. coli BL21(DE3) using pET26b(+),pET26b(+), and the recombinant MAP was purified by using an Ni-NTA column. The size of recombinant MAP was 29 kDa as determined by SDS-PAGE. The optimum pH and temperature of CY54 MAP were pH 5.0 and 60℃, respectively. The activity of CY54 MAP was most significantly increased by Co²⁺ ion (159%), and showed the highest activity at 12% NaCl. K_m and V_max were 0.64 ± 0.006 mM and 10.12 ± 0.014 U/mg protein, respectively when met-pNA was used as the substrate. This is the first report on a MAP from Tetragenococcus species.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2003.10a
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• pp.170-177
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• 2003

Gene expression data are the quantitative measurements of expression levels and ratios of numberous genes in different situations based on microarray image analysis results. The process to draw meaningful information related to genomic diseases and various biological activities from gene expression data is known as gene expression data analysis. In this paper, we present a hierarchical clustering method of gene expression data based on self organizing map which can analyze the clustering result of gene expression data more efficiently. Using our proposed method, we could eliminate the uncertainty of cluster boundary which is the inherited disadvantage of self organizing map and use the visualization function of hierarchical clustering. And, we could process massive data using fast processing speed of self organizing map and interpret the clustering result of self organizing map more efficiently and user-friendly. To verify the efficiency of our proposed algorithm, we performed tests with following 3 data sets, animal feature data set, yeast gene expression data and leukemia gene expression data set. The result demonstrated the feasibility and utility of the proposed clustering algorithm.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 1997.06a
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• pp.23-49
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• 1997

This is a progress report of rice biotechnology including development of gene transformation system, gene cloning and molecular mapping in rice. The scope of the research was focused on the connection between conventional breeding and biotech-researches. Plant transformation via Agrobacterium or particle bombardment was developed to introduce one or several genes to recommended rice cultivars. Two chimeric genes containing a maize ribosome inactivating protein gene (RIP) and a gerbicide resistant gene (bar) were introduced to Nipponbare, a Japonica cultivar, and transmitted to Korean cultivars. The homozygous progenies of herbicide resistant transgenic plant showed good fertility and agronomic characters. To explore the genetic resourses in rice, over 8,000 cDNA clones from immature rice seed have been isolated and sequenced. About 13% of clones were identified as enzymes related to metabolic pathway. Among them, twenty clones have high homology with genes encoding enzymes in the photorespiratory carbon cycle reaction. Up to now about 100 clones were fully sequenced and registered at EMBL and GenBank. For the mapping of quantitative tarits loci (QTL) and eternal recombinant inbred population with 164 F13 lines (MGRI) was developed from a cross between Milyang 23 and Gihobyeo, Korean rice cultivars. After construction of fully saturated RFLP and AFLP map, quantitative traits using MGRI population were analyzed and integrated into the molecular map. Eighty seven loci were determined with 27 QTL characters including yield and yield components on rice chromosomes. Map based cloning was also tried to isolate semi-dwarf (sd-1) gene in rice. A DNA probe, RG 109, the most tightly linked to sd-1 gene was used to screen from bacterial artifical chromosome (BAC) libraries and five over lapping clones presumably containing sd-1 gene were isolated. Rice genetic database including results of biotech reasearch and classical genetics is provided at Korea Rice Genome Server which is accessible with world wide web (www) browser. The server provides rice cDNA sequences and map informations linked with phenotypic images.

• Proceedings of the KSAR Conference
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• 2001.10a
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• pp.14-17
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• 2001

Positional cloning (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150kb of DNA was identified. A gene associated with this deletion was identified using cDNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

• The Journal of Korean Obstetrics and Gynecology
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• v.15 no.4
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• pp.1-16
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• 2002

This work examines the effect of treatment with Gyukhachukeotang on the growth of uterine myomal cells. Comparisons of cell growth, MAP kinase activity and expression of bcl-2 (apoptosis-related gene) were made between the control and experimental samples. The results as fallows; 1. Any concentration of Gyukhachukeotang above 0.01% yielded growth inhibition. Concentrations of 5% and 10% stopped all cell growth, demonstrating the effectiveness of Gyukhachukeotang as a growth inhibitor on uterine myomal cells. 2. The MAP kinase activity in uterine myomal cells treated with Gyukhachukeotang was decreased to a high degree at the concentration of 10%, and some inhibition of activity was detected at a concentration of 5%. 3. The expression of bcl-2, a Cell Apoptosis-related gene, in uterine myoma cells treated with Gyukhachukeotang was gradually increased with increasing concentration of Gyukhachukeotang. These results indicate the ability of Gyukhachukeotang to control uterine myomal cell growth, with concurrent reduction of MAP kinase activity. Treatment with Gyukhachukeotang appears to trigger a normal apoptosis response, as indicated by increased bcl-2 expression. This observed increase in apoptosis indicates that Gyukhachukeotang is an appropriate prescription to treat uterine myomal cells.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.10a
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• pp.14-17
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• 2001

Positional clonging (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150 kb of DNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

• Korean Journal of Microbiology
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• v.28 no.1
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• pp.27-34
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• 1990

Two macrolide-lincosamide-streptogramin B (MLS) antibiotic resistance genes, one expressed inducibly and the other expressed constitutively were recognized from a single Staphylococcus aureus DH1 strain. The inducible MLS resistance gene was isolated and cloned from the R-plasmid pDE1(7.4kb) and the constitutive gene was from chromosomal DNA. Base sequence of the inducible MLS resistance gene (1.2kb) was determined and found as same that of pE194. The restriction map of the cloned constitutive MLS resistance gene was compared with that of the inducible gene. Two genes have same restriction map except leader region. In the constitutive gene there is no leader region which is doing major role in inducible expression.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.255-267
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• 2015

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic debilitating disease affecting ruminants worldwide. In the present study, we aimed to determine the major gene networks and pathways underlying the immune response to MAP infection using whole-blood cells, as well as provide the potential transcriptional markers for identifying the status of MAP infection. We analyzed the transcriptional profiles of whole-blood cells of cattle identified and grouped according to the presence of MAP-specific antibodies and the MAP shed by them. The grouping was based on the results obtained by ELISA and PCR analyses as follows: i) Test1 group: MAP-negative results obtained by ELISA and positive results obtained by PCR; ii) Test2 group: MAP-positive results obtained by ELISA and negative results obtained by PCR; iii) Test3 group: MAP-positive results obtained by ELISA and positive results obtained by PCR; iv) uninfected control: MAP-negative results obtained both by ELISA and PCR analysis. The results showed down-regulated production and metabolism of reactive oxygen species in the Test1 group, activation of pathways related to the host-defense response against MAP (LXR/RXR activation and complement system) in the Test2 and Test3 groups, and anti-inflammatory response (activation of IL-10 signaling pathway) only in the Test3 group. Our data indicate a balanced response that serves the immune-limiting mechanism while the host-defense responses are progressing.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.297-302
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• 1985

The 7.1 Mdal Xbaf fragment of Bacillus pasteurii ATCC 11859 containing gene for urease was inserted into the Xbal site of bifunctional plasmid pGR71, and its urease gene was cloned and expressed in E. coil RRI. But the cloned gene was not expressed in Bacillus subtilis BR151 in consequence of deletion of inserted DNA fragment. The recombinant plasmid thus formed was named pGU66. The restriction map of the plasmid pGU66 was determined, and the size of the plasmid was estimated to be 12.6 Mdal by double digestion of restriction enzymes of the plasmid. The urease of the cloned strain was accumulated in periplasmic space and very similiar to that of donor strains in their enzymatic properties.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.167-172
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• 1988

A genetic map of the IncP-1 group plasmid pKU10 has been prepared through the construction of recombinant plasmids containing various fragments of pKU10. Phenotypic analysis of these derivatives has identified the location of genes encoding resistance to ampicillin, tetracyclin, and chloramphenicol. The region involved in conferring resistance to ampicillin was located around two PstI sites that are 1.0Kb apart. The tetracyclin resistance gene was mapped on the region of HindIII E fragment and a part of HindIII D fragment, and the determinant for chloramphenicol resistance gene was localized on HindIII D fragment.