• KSBB Journal
• /
• v.11 no.4
• /
• pp.431-437
• /
• 1996

The nar promoter of Escherichta coli, which is maximally induced under anaerobic conditions in the presence of nitrate, was characterized to see whether the nar promoter cloned onto pBR322 can be used as an expression promoter. The modified nar promoter, in which several bases in the -10 region was mutated to the consensus sequence by site-directed mutagenesis, was characterized in E. coli, on which chromosome the fnr gene affecting expression of the nar promoter according to dissolved oxygen level was mutated. The E. coli lacZ gene was used as a reporter gene. The following effects were investigated to find optimal conditions to induce the modified nar promoter: induction methods, optimal nitrate concentrations, the amount of ${\beta}$-galactosidase expressed at the different growth conditions, and induction characteristics. The following results were obtained from the experiments : expression of ${\beta}$-galactosidase from the modified nar promoter was not affected much by nitrate concentrations. The maximal specific ${\beta}$-galactosidase activity was obtained when E. coli was grown under aerobic conditions, and then the modified nar promoter was induced at OD600=2.2 under microaerobic conditions (DO=1∼2%), under which conditions the maximal specific ${\beta}$-galactosidase activity was 13,000 Miller units. However, the specific ${\beta}$-galactosidase activity was approximately 6,000 Miller units even before the modified nar promoter was induced. Therefore, the modified nar promoter seemed to be useful when the cloned gene wants to be expressed in E. coli constitutively.

• Journal of Aquaculture
• /
• v.13 no.4
• /
• pp.331-338
• /
• 2000

To establish an alternate technology for all male seed production of Nile tilapia, Oreochromis niloticus at hatchery scale, the performance of progenies produced by crossing superfemale with sex reversed male was evaluated. Free crossings between 21 superfemales and 7 sex reversed males stocked in an aquarium for 60 days resulted in the production of 93.5 % male progenies, compared to 53.2 % males produced by an equal number of normal females and normal males. Spawning frequency was lower (19 times) in the experimental cross including superfemales than that (24 times) in the control including normal females. Seed production in the experimental aquarium was (3,085) significantly lower than that (3,797) of the control. Both in terms of seeds per female per day or seeds per g female per day, seed productivity did not significantly differ between these groups. The alternate technology involving crossing between superfemale and sex reversed male has proven to be an efficient alternate technology for mass production of all male seeds at the hatchery scale.

• Journal of Aquaculture
• /
• v.13 no.4
• /
• pp.325-330
• /
• 2000

For development of all male seed production at hatchery scale, a comparative study was made on the seed production traits of supermale (IT - t) and superfemale (IT - L1 ~) Nile tilapia, Oreochromis niloticus. Supermales were crossed with normal females (XX- f) and the superfemales with normal males (XY - t ) or sex reversed males (XX- L1 t); progeny survival of these crossings and sex ratio were evaluated. Hatching success of the eggs, fertilized by the supermale was significantly lower than that by the normal male. Over 95-99 % progenies sired by crossing supermales with normal females were males, while 52-55 % progenies alone were males with the cross of normal males and normal females. Hatching success and survival of alevins were significantly higher for the progenies of the crosses between superfemales and sex reversed males than those resulting from the crosses between superfemales and normal males. However, there was no significant difference in the sex ratio among the progenies of these crosses. Therefore, crossings of superfemales with sex reversed males provide the highest percentage of survival and male progenies.

• Journal of Korean Biological Nursing Science
• /
• v.5 no.1
• /
• pp.13-22
• /
• 2003

The purpose of this study was to investigate and analyze current educational requirements related to genetics curriculum(from June 2002 to September 2002) established at nursing institutions and to provide the basic data for the development of genetics science program at the undergraduate. Subjects of this study were comprised of twenty-three colleges of nursing in 4-year baccalaureate and thirty colleges in 3-year diploma programs. The results of this study were as follows : 1) 32 colleges offer courses related to genetics. 29 among 32 colleges have that integrated. Three schools have established completely independent courses of genetics. 21 colleges do not have any courses dealing with genetics. 2) The contents of courses related to genetics include: Congenital abnormalities, chromosomal aberrations, congenital metabolic disease, prenatal diagnosis and genetic counseling, genes and chromosomes, immune genetics, blood type and genetics, rule of genetics, variation in gene expression, the map of the human gene, gene linkage genetics, interaction of genes, single inheritance in order and genetic biochemistry. 3) For course credit, 14colleges(48.3%) offered at most 1 credit per course. The grade of student who can take the course, 51.7% were in their second year while 37.9% were in their third year. The majors of nursing faculty who taught the course were nursing(51.7%) and basic nursing science(17.2%). 4) As far as the need of opening the courses related to genetics, 36 colleges(67.0%) have made a 'need', 12 schools(22.6%) state 'dose not need'. 711e reason for need were the following development of bio engineering, increase number of patients who are related to genetics, recognition of the need in clinical nursing. 7 schools(13.2%) agreed to offer independent course in genetics but 39 schools(73.6%) are in disagreement with that. When the school offers the course with other courses, 27 schools(50.0%) are opening basic nursing science and 14 schools(26.4%) are opening nursing as an integrated courses. If the name of course was either genetic nursing(34.0%) or genetics(28.3%), the credits for the course was one or 2 credits. 33 schools(62.3%) students were in the first or second years. 41 schools(84.9%), the majors of the faculty who had taught the course were either basic nursing science(35.8%), nursing(28.3%) or basic medicine(24.5%). The contents of the course should include in that order: Chromosome aberrations, prenatal diagnosis and genetic counseling, congenital metabolic disease, congenital abnormalities, genes and chromosomes, the rules of genetics, immune genetics, interaction of genes, variation in gene expression, etc. The results and discussions of the study indicate that the entire curriculums need to be investigated with respect to contents of education, nursing curriculums and name of courses because of the increasing need of knowledge related to genetics in the clinical practice.

• Proceedings of the Korean Society for Bioinformatics Conference
• /
• 2001.10a
• /
• pp.61-86
• /
• 2001

All cancers are caused by abnormalities in DNA sequence. Throughout life, the DNA in human cells is exposed to mutagens and suffers mistakes in replication, resulting in progressive, subtle changes in the DNA sequence in each cell. Since the development of conventional and molecular cytogenetic methods to the analysis of chromosomal aberrations in cancers, more than 1,800 recurring chromosomal breakpoints have been identified. These breakpoints and regions of nonrandom copy number changes typically point to the location of genes involved in cancer initiation and progression. With the introduction of molecular cytogenetic methodologies based on fluorescence in situ hybridization (FISH), namely, comparative genomic hybridization (CGH) and multicolor FISH (m-FISH) in carcinomas become susceptible to analysis. Conventional CGH has been widely applied for the detection of genomic imbalances in tumor cells, and used normal metaphase chromosomes as targets for the mapping of copy number changes. However, this limits the mapping of such imbalances to the resolution limit of metaphase chromosomes (usually 10 to 20 Mb). Efforts to increase this resolution have led to the "new"concept of genomic DNA chip (1 to 2 Mb), whereby the chromosomal target is replaced with cloned DNA immobilized on such as glass slides. The resulting resolution then depends on the size of the immobilized DNA fragments. We have completed the first draft of its Korean Genome Project. The project proceeded by end sequencing inserts from a library of 96,768 bacterial artificial chromosomes (BACs) containing genomic DNA fragments from Korean ethnicity. The sequenced BAC ends were then compared to the Human Genome Project′s publicly available sequence database and aligned according to known cancer gene sequences. These BAC clones were biotinylated by nick translation, hybridized to cytogenetic preparations of metaphase cells, and detected with fluorescein-conjugated avidin. Only locations of unique or low-copy Portions of the clone are identified, because high-copy interspersed repetitive sequences in the probe were suppressed by the addition of unlabelled Cotl DNA. Banding patterns were produced using DAPI. By this means, every BAC fragment has been matched to its appropriate chromosomal location. We have placed 86 (156 BAC clones) cytogenetically defined landmarks to help with the characterization of known cancer genes. Microarray techniques would be applied in CGH by replacement of metaphase chromosome to arrayed BAC confirming in oncogene and tumor suppressor gene: and an array BAC clones from the collection is used to perform a genome-wide scan for segmental aneuploidy by array-CGH. Therefore, the genomic DNA chip (arrayed BAC) will be undoubtedly provide accurate diagnosis of deletions, duplication, insertions and rearrangements of genomic material related to various human phenotypes, including neoplasias. And our tumor markers based on genetic abnormalities of cancer would be identified and contribute to the screening of the stage of cancers and/or hereditary diseases

• Journal of Genetic Medicine
• /
• v.2 no.2
• /
• pp.53-57
• /
• 1998

Spinal muscular atrophy (SMA) type I is a common severe autosomal recessive inherited neuromuscular disorder that has been mapped to chromosome 5q11.2-13.3. The survival motor neuron (SMN) gene, a candidate gene, is known to be deleted in 96% of patients with SMA type I. Presently, PCR and single strand conformation polymorphism (PCR-SSCP) analyses have been made possible for application to both archival slides and paraffin-embedded tissues. Archival materials represent valuable DNA resources for genetic diagnosis. We applied these methods for the identification of SMN gene of SMA type I in archival specimens for the prenatal diagnosis. In this study, we performed the prenatal diagnosis with chorionic villus sampling (CVS) cells on two women who had experienced neonatal death of SMA type I. DNA extraction was done from archival slide and tissue materials and PEP-PCR was performed using CVS cells. In order to identify common deletion region of SMN and neuronal apoptosis-inhibitory protein (NAIP) genes, cold PCR-SSCP and PCR-restriction site assay were carried out. Case 1 had deletions of the exons 7 and 8, and case 2 had exon 7 only on the telomeric SMN gene. Both cases were found to be normal on NAIP gene. These results were the same for both CVS and archival biopsied specimens. In both cases, the fetuses were, therefore, predicted to be at very high risk of being affected and the pregnancy were terminated. These data clearly demonstrate that archival slide and paraffin-embedded tissues can be a valuable source of DNA when the prenatal genetic diagnosis is needed in case any source for genetic analysis is not readily available due to previous death of the fetus or neonate.

• Journal of Genetic Medicine
• /
• v.5 no.2
• /
• pp.125-130
• /
• 2008

Propose: To analyze the indications and cytogenetic results of midtrimester amniocentesis. Material and Methods: This study reviewed 2,523 cases of midtrimester prenatal genetic amniocentesis performed at MizMedi Hospital between January 2000 and December 2007. Results: The most frequent indication for midtrimester amniocentesis was advanced maternal age (45.9%), followed by positive serum markers (29.9%). Chromosomal aberrations were diagnosed in 110 cases (4.4%), for which numerical aberration accounted for 38 cases (34.5%), structural aberration accounted for 65 cases (59.1%), and mosaicism accounted for 7 cases (6.4%). Among the autosomal aberrations, there were 20 cases of trisomy 21 and 8 cases of trisomy 18. With respect to structural aberrations, there were 14 cases of reciprocal translocation and 8 cases of robertsonian translocation. The frequencies of chromosomal aberrations according to the indication were highest in individuals with a family history of chromosome abnormality 14.0% (8/57) followed by previous congenital anomaly 5.9% (2/34). Conclusion: Midtrimester amniocentesis is an effective tool for prenatal diagnosis. Indications such as advanced maternal age, maternal serum markers, and ultrasound are important for predicting abnormal fetal karyotypes.

• Clinical and Experimental Pediatrics
• /
• v.54 no.8
• /
• pp.335-339
• /
• 2011

Purpose: Hyperhomocysteinemia is known as a risk factor for atherosclerosis. Preclinical arteriosclerosis is noted and premature atherosclerosis is known to be accelerated in Kawasaki disease (KD) patients. Genetic polymorphisms in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene result in elevated plasma homocysteine concentrations and are known to be associated with the development of coronary artery disease. Our hypothesis is that single nucleotide polymorphisms (SNPs) of the MTHFR gene are related to the development of KD and coronary artery lesions (CALs). Methods: For this study, we selected 3 candidate single nucleotide polymorphisms (SNPs) (rs2274976, rs1801131, and rs1801133) of MTHFR. These SNPs are located on chromosome 1p36.3. We included 101 KD patients and 306 healthy adults as controls in this study. CALs were seen in 38 patients. Genotypes of the selected SNPs were determined by direct sequencing and analyzed with SNPAlyze. Results: The genetic distribution and allelic frequency of the 3 MTHFR SNPs (rs2274976, rs1801131, and rs1801133) were not significantly different in patients with KD compared to the control group (P=0.71, 0.17, and 0.96, respectively). There was no difference in the genetic distribution of the MTHFR SNPs between the normal control group and the CAL group (P=0.43, 0.39, 0.52 respectively). Conclusion: The genetic distribution of the MTHFR SNPs (rs2274976, rs1801131, and rs1801133) was not different in the KD group compared to the control group. In addition, the genetic distribution of these SNPs was not different in the CAL group compared to the control group in the Korean population.

• Biomedical Science Letters
• /
• v.26 no.3
• /
• pp.230-237
• /
• 2020

Staphylococcus aureus (S. aureus) is known as a bacterium that can cause skin infections, respiratory system infections, and sinusitis; however, it can exist as a normal flora rather than a pathogen. Recently, methicillin-resistant S. aureus (MRSA) infections have emerged in the community as a new variant of community-associated (CA)-MRSA. In the present study, S. aureus and MRSA were isolated and cultured by collecting samples from facilities and environments where students and educational personnel have multiple contacts on university campuses; specifically, the nostrils and hands of college students were tested from July to September of 2019. The molecular properties of the isolated MRSA were analyzed, and the one MRSA strain was isolated from the university campuses. One MRSA that was isolated and cultured on campus was the mec complex group A and staphylococcal cassette chromosome (SCC) mec type II, which is a characteristic of healthcare-associated (HA)-MRSA, and SCCmec type V, which is a characteristic of CA-MRSA. This result was similar to other studies wherein the SCCmec type II was detected in SCCmec typing analysis in CA-MRSA. To confirm whether there is a new variant of CA-MRSA in the Republic of Korea, additional follow-up studies on the analysis of virulence factors of MRSA are needed by additionally separating CA-MRSA from the body parts of university students and educational personnel.

• Molecular & Cellular Toxicology
• /
• v.5 no.2
• /
• pp.126-132
• /
• 2009

The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.