• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.176-181
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• 1999

The variations of gene expression and pituitary contents of GTH subunits during the ovarian development of masu salmon, Oncorhynchus masou, were investigated. The pituitary GTHs contents was measured by radioimmunoassays (RIAs) using purified GTH subunits and their antibodies. Pituitary contents of GTH $I\beta$ gradually increased from April through August, and reached the maximum in October. On the other hand, pituitary contents of GTH $II\beta$ remained low until August, but they rapidly increased in October. Total RNAs were prepared from pooled pituitaries and the GTH subunits mRNA in pituitaries was quantified by Northern blot hybridization using masu salmon cDNA probes for each GTH subunit. GTH $I\beta$ mRNA level increased with the progression of ovarian maturity. However, GTH $II\beta$ mRNA was detected only at a more advanced stage, and were extremly high at ovulation. A high levels for GTH a mRNA was detected only at ovulation stage. The synchronous increase in pituitary contents and mRNA levels suggested that ovarian maturity in masu salmon was regulated by both GTH I and GTH II.

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.39-46
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• 2001

To examine the effect of copy number-dependent transgenic genotype on the expression of foreign gene, stable hemizygous and homozygous transgenic breeding line was established using artificial parthenogenesis. For this purpose, induced diploid gynogenetic transgenesis was optimized in mud loach (Misgurnus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to $4,050\;ergs/mm^2$ The UV-irradiated sperm were inseminated into eggs from recessive color strain (yellow) or heterozygous transgenic mud loach containing CAT gene. Cold shock at $2^{\circ}C$ for 60 min, 5 min post fertilization successfully restored the diploidy of eggs inseminated with UV-irradiated sperm. Restoration to diploidy was confirmed by flow cytometry and gynogenetic status was verified by examining maternal exclusive inheritance of multi-locus DNA fingerprints, body color and transgenic marker. Putative isogenic transgenic fish clearly showed homozygous status at trans gene locus based on Southern blot hybridization and progeny testing. Further, such homozygous gynogenetic diploids revealed the increased levels of transgene expression, when compared to those of heterozygous (hemizygous) transgenic fish.

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.207-216
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• 2018

Solanum berthaultii is one of the wild diploid Solanum species, which is an excellent resource in potato breeding owing to its resistance to several important pathogens. On the other hand, sexual hybridization between S. berthaultii and S. tuberosum (potato) is limited because of their sexual incompatibility. Therefore, cell fusion can be used to introgress various novel traits from this wild species into the cultivated potatoes. After cell fusion, it is crucial to identify fusion products with the aid of molecular markers. In this study, the chloroplast genome sequence of S. berthaultii obtained by next-generation sequencing technology was described and compared with those of five other Solanum species to develop S. berthaultii specific markers. A total sequence length of the chloroplast genome is 155,533 bp. The structural organization of the chloroplast genome is similar to those of the five other Solanum species. Phylogenic analysis with 25 other Solanaceae species revealed that S. berthaultii is most closely located with S. tuberosum. Additional comparison of the chloroplast genome sequence with those of the five Solanum species revealed 25 SNPs specific to S. berthaultii. Based on these SNPs, six PCR-based markers for differentiating S. berthaultii from other Solanum species were developed. These markers will facilitate the selection of fusion products and accelerate potato breeding using S. berthaultii.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7731-7735
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• 2014

Colorectal cancer (CRC) is the third most common cause of death due to cancer in the worldwide and the incidence is also increasing in Turkey. Our present aim was to investigate any association between germ-line methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms and CRC risk in Turkey. A total of 86 CRC cases and 212 control individuals of the same ethnicity were included in the current study. Peripheral blood-DNA samples were used for genotyping by StripAssay technique, based on the reverse-hybridization principle and real-time PCR methods. Results were compared in Pearson Chi-square and multiple logistic regression models. The MTHFR 677TT (homozygous) genotype was found in 20.9% and the T allele frequency 4.2-fold increased in CRC when compared with the control group.The second SNP MTHFR 1298CC (homozygous) genotype was found in 14.0% and the C allele frequency 1.4-fold elevated in the CRC group. The current data suggest strong associations between both SNPs of germ-line MTHFR 677 C>T and 1298 A>C genotypes and CRC susceptibility in the Turkish population. Now the results need to be confirmed with a larger sample size.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.29-29
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• 2003

Mycoplasma hyopneumoniae is among the most prevalent and important infectious agents associated with porcine respiratory disease complex. The airway dagame caused by M. hyopneumoniae adversely affect the pulmonary host defense mechanisms and may lead to secondary bacterial infections. Culture is considered to be the "gold standard" for diagnosis but this is a very slow and labor-intensive procedure. Isolation of M. hyopneumoniae is complicated by its fastidious nature and extremely slow growth. Thirty days of incubation may be necessary to detect the organism in primary broth cultures. The purposes of the study were (ⅰ) to develop nested PCR for the detection of M. hyopneumoniae for the detection of M. hyopneumoniae DNA in the formalin-fixed, paraffin-embedded lung tissues from experimentally and naturally infected pigs and (ⅱ) to compare the utility of nested PCR with in situ hybridization. (omitted)

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2003.09a
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• pp.152-157
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• 2003

Diesel-contaminated soils were ozonated for different times (0 - 900 min) and incubated for 9 wk to monitor petroleum hydrocarbons (PH)-degradative potential of indigenous microorganisms in the soils. Increased ozonation time decreased not only concentration of PH but also number of microorganisms in the soils. Microorganisms in the ozonated soils increased during 9-wk incubation as monitored by culture- and nonculture-based methods. Higher (1-2 orders of magnitude) cell number was observed by quantitative analysis of soil DNA using probes detecting genes encoding 165 rRNA(rrn), naphthalene dioxygenase (nahA), toluene dioxygenase (todC), and alkane hydroxylase (alkB) than microbial abundance estimated by culture-based methods. Such PH-degraders were relatively a few or under detection limit in 900-min ozonated soil. Further PH-removal observed during the incubation period supported the presence of PH-degraders in ozonated soils. Highest reduction (25.4%) of total PH (TPH) was observed in 180-min ozonated soil white negligible reduction was shown in 900-min ozonated soil during the period, resulting in lowest TPH-concentration in 180-min ozonated soil among the ozonated soils. Microbial community composition in 9-wk incubated soils revealed slight difference between 900-min ozonated and unozonated soils as analyzed by whole cell hybridization using group-specific rRNA-targeted oligonucleotides. Results of this study suggest that appropriate ozonation and subsequent biodegradation by indigenous microorganisms may be a cost-effective and successful remediation strategy for PH-contaminated soils.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.1
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• pp.94-106
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• 1990

사람들이나 동물의 소화관의 상재균으로서 중요한 Lactobacillus acidophilus의 여러균주를 사용하여 세포표면의 성상을 조사하였다. 불리한 세포벽의 negative 염색한 표본을 전자현미경으로 검토한 결과 사용한 21주중 15주는 세포벽외층에 단백질로 구성딘 regular array를 생산하고 있음을 알았다. RA를 구성하는 단백질의 분자량은 41KDa-48KDa이었다 아미노사노성 및 Staphylococcus au-reus V8 protease로 한정분해 후 및 N-chlorosuccini-mide로 부분절단후의 peptide map에서 Johnson등의 분류의 subgroup A-1에 속하는 균주가 생산하는 RA단백질은 불균질하였으나 그외 균주의 RA단백질은 같은 subgroup에 속하고 있어도 불균질하였음을 알았다 따라서 RA단백질은 불균질하였으나 그외 균주의 RA단백질은 같은 subgroup에 속하고 있어도 불균질하였음을 알았다 따라서 RA생산의 유무 및 RA단백질의 분자량측정과 peptide mapping을 행하면 지금까지 행해온 DNA-hybridization과 같은 복잡한 절차를 사용하지 않아도 subgroup A-1의 L. acidophilus가 동정 가능함을 알았다 L acidophilus의 세포표면소수성은 일반적으로 RA를 생산하지 않는 균주보다는 RA를 생사하는 균주에 높은 것이많았다 RA생산균주에 있어서는 RA단백질은 세포표면소수성과 직접관계하고 있는 것은 적었다.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.177-177
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• 2003

Methylmercury (MeHg) is a well-known neurotoxicant that causes severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, suppressive subtractive hybridization (SSH) was performed to identify differentially expressed genes on human neuroblastoma cell line, SH-SY5Y treated with DMSO and MeHg (6.25 uM) for 6 hr. Differentially expressed cDNA clones were sequenced and were screened by dot blot to eliminate false positive clones. 13 of 35 screened genes were confirmed using real time RT-PCR. These genes include EB1,90-kDa heat-shock protein, chromosome condensation-related SMC-associated protein and brain peptide Al, etc. Analysis of these genes may provide an insight into the neurotoxic effects of MeHg in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.716-721
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• 2002

The ldhL gene encoding the $_L$-(+) lactate dehydrogenase was cloned from Lactobacillus reuteri ATCC 55739 chromosomal DNA and characterized. An internal 750-bp fiagment of ldhL gene was amplified by PCR using primers based on the conserved region of lactobacilli ldhL genes. A genomic library off. reuteri ATCC 55739 was constructed using pBR322, and colony hybridization experiments were performed using the 750-bp fragment as aprobe. One clone harboring a 4.0-kb PstI fragment was identified, and nucleotide sequencing confirmed it as an open reading frame of 972 bp in size in the middle. In addition to IdhL gene, an ORF homologous to Streptococcus pneumoniae TIGR4 hydrolase gene and 3' part of phosphomevalonate kinase gene (mvaK2) were also found on the 4 kb fragment. $_L$-LDH of L. reuteri ATCC 55739 showed the highest degree of homology with the $_L$-LDH of Pediococcus acidilactici (62.4%), fullowed by the $_L$-LDH of Lactobacillus pentosus (58.7%). The size of IdhL transcript determined by Northern blot was 1 kb, indicating the monocistronic nature of IdhL.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1995.06b
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• pp.1-19
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• 1995

The plasmid pJEL 101 contains a highly repetitive element from the genome of Xanthomonas oryae pv. oryzae that has properties of an insertional element. The insertional nature of the element, hereto referred to as IS203, was confirmed by molecular analyses of the element and three related elements that were isolated from X. oryzae. The related sequences were isolated on the basis of transposition to the transposon-trapping vector pL3SAC and hybridization with pJEL101. The trapped elements (IS203a, IS203b, and IS203c) were each composed of 1,055 base pairs with 25 base terminal inverted repeats. The elements caused a three base pair target site duplication at the site of insertion in the sacRB gene. The sequence of pJEL 101 has 96% base pair identity with IS203a and 99% identity with IS203a and IS203c but lacks three nucleotides of the consensus left terminal repeat. IS203b has the same DNA sequences as IS203c but is inserted ito the sacRB gene in the opposite orientation. The longest open reading frame of IS203a could code for a protein of 318 amino acids and molecular weight of 37, 151. A search of the Genbank database revealed that IS203 has 51% identity with 909 nucleotides of IS4551 from Escherichia coli. The predicted protein of ORF1 has 40% and 30% amino acid identity to the ORF1 of Tn4551 and the transposase of IS30, respectively.