• Radiation Oncology Journal
• /
• v.23 no.1
• /
• pp.43-50
• /
• 2005

Purpose : A number of genes and their products are Induced early or late following exposure of cells to ionizing radiation. These radiation-Induced genes have various effects on irradiated cells and tissues. Suppression subtractive hybridization (SSH) based on PCR was used to Identify the differentially expressed genes by radiation in cervix carcinoma cells. Materials and Methods : Total RNA and poly $(A)^+$ mRNA were Isolated from Irradiated and non-irradiated HeLa cells. Forward- and reverse-subtracted cDNA libraries were constructed using SSH. Eighty-eight clones of each were used to randomly select differentially expressed genes using reverse Northern blotting (dot blot analysis). Northern blotting was used to verify the screened genes. Results : Of the 17t clones, 10 genes in the forward-subtracted library and 9 genes In the reverse-subtracted library were identified as differentially expressed radiation-induced genes by PCR-select differential screening. Three clones from the forward-subtracted library were confirmed by Northern blotting, and showed increased expression in a dose-dependent manner, including a telomerase catalytic subunit and sodium channel-like protein gene, and an ESTs (expressed sequence tags) gene. Conclusion : We Identified differentially expressed radiation-induced genes with low-abundance genes with SSH, but further characterization of theses genes are necessary to clarify the biological functions of them.

• Journal of Marine Bioscience and Biotechnology
• /
• v.1 no.3
• /
• pp.186-192
• /
• 2006

Short interspersed repetitive elements (SINEs) are dispersed throughout eukaryotic genomes. These SINEs have been shown to be excellent phylogenetic markers for the closed related species. In this report, we isolated two novel families of SINEs from the mud loach. The two SINE families, mlSINE-L and mlSINE-S, have genomic lengths of about 410bp and 270bp, respectively. 5' and 3' ends of the SINE families are well conserved and highly homologous to each of corresponding ends of RSg-1 and SmaI SINEs. Phylogenetic analysis shows that mlSINEs are unique to the mud loach. A dot blot hybridization experiment shows that mlSINE-L has an estimated copy number of $1{\times}10^3$ per $2{\times}10^9bp$ (2.8 pg) and is more frequently distributed at nuclear matrix attachment regions (MARs) than loop DNAs. The result suggests that mlSINEs may preferentially integrate in or near MARs.

• Journal of Korean Society of Occupational and Environmental Hygiene
• /
• v.21 no.1
• /
• pp.49-54
• /
• 2011

In order to develop the methods for exposure assessment and find susceptibility markers for the workers who are exposed to low doses of toluene, xylene and other chemical in petroleum industries, we investigated the application of P-450 expression in human lymphocytes utilizing mouse monoclonal anti-rat CYP2B1/2, the levels of toluene and xylene in air and their metabolite levels in urine with the levels of expressed CYP2B1/2 proteins. The general characteristics such as age, smoking and drinking habit were no significant difference between the control and exposed workers, but the working durations and working hours were significantly different. Workers in exposed group were exposed to the mean of 2.1 ppm (range, 0.00-4.54) of toluene and 0.3 ppm (rang, 0.00-1.23) of xylene. The mean concentration of urinary hippuric acid was low and less than 1/5 of the biological exposure index recommended by the Ministry of Employment and Labor Korea. Methyl hippuric acid in urine was not detected in control and exposed workers. Also, there were no significant differences in the levels of the urinary metabolites between the control and exposed group. When chemiluminescence dot blottings were carried out utilizing mouse monoclonal antibody against CYP2B1/2, the strong density dots corresponding to a mouse monoclonal antibody was observed in the human lymphocytes from the exposed workers. These results suggested that the chemiluminescence dot blot assay for CYP of lymphocytes should be valuable for identifying CYP expression as biomarkers in the workers exposed to toluene and xylene.

• The Korean Journal of Mycology
• /
• v.27 no.1 s.88
• /
• pp.20-26
• /
• 1999

The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

• Korean Journal of Veterinary Research
• /
• v.43 no.4
• /
• pp.667-675
• /
• 2003

The safety of Brucella abortus strain RB51(SRB51) was investigated in dairy cows and Korean native cattle of 4~7th month of gestation. From experimentally inoculated cattle, 18 of 25 (72.0%) dairy cows, and 3 of 10 (30.0%) Korean native cattle were aborted or delivered premature fetus. There were no significant differences in the incidence of abortion depending on the inoculation route (intramuscular and subcutaneous) and dosages ($1{\times}10^9$, $2.8{\times}10^9$, and $4.0{\times}10^9$ CFU). The antibodies to the SRB51 were measured by a dot blot enzyme-linked immunosorbent assay. The highest titers to SRB51 were detected between 5~7 weeks after inoculation and the specitic antibody could be detected up to 28 weeks after inoculation. The SRB51 was isolated from amnio-allantoic fluid, bronchial lymph node, mammary gland, and supramammary lymph node in 5 of 25 dairy cows during 4 weeks after either abortion or delivery. Although SRB51 was isolated from 4 of 24 aborted fetus or normally delivered calves at parturition time, it was not isolated during 4 weeks afterward. Eleven of twentyfive dairy cows showed the endometritis and/or necrosis until 6 weeks after delivery, no lesions were seen at 8 weeks after delivery and uterus from control dairy cows. The results of present study revealed that SRB51 might induce the clinical signs of brucellosis in the pregnant cattle at 4~7th month of gestation.

• BMB Reports
• /
• v.35 no.4
• /
• pp.420-427
• /
• 2002

Partial nerve injury is the main cause of neuropathic pain disorders in humans. Acupuncture has long been used to relieve pain. It is known to relieve pain by controlling the activities of the autonomic nervous system. Although the mechanism of neuropathic pain and analgesic effects of electroacupuncture (EA) have been studied in a rat model system, its detailed mechanism at the molecular level remains unclear. To identify genes that might serve as either markers or explain these distinct biological functions, a cDNA microarray analysis was used to compare the expression of 8,400 genes among three sample groups. Messenger RNAs that were pooled from the spinal nerves of 7 normal. 7 neuropathic pain, and 7 EA treatment rat models were compared. Sixty-eight genes were differentially expressed more than 2-fold in the neuropathic rat model when compared to the normal, and restored to the normal expression level after the EA treatment. These genes are involved in a number of biological processes, including the signal transduction, gene expression, and nociceptive pathways. Confirmation of the differential gene expression was performed by a dot-blot analysis. Dot-blotting results showed that the opioid receptor sigma was among those genes. This indicates that opioid-signaling events are involved in neuropathic pain and the analgesic effects of EA. The potential application of these data include the identification and characterization of signaling pathways that are involved in the EA treatment, studies on the role of the opioid receptor in neuropathic pain, and further exploration on the role of selected identified genes in animal models.

• Food Science of Animal Resources
• /
• v.34 no.3
• /
• pp.339-345
• /
• 2014

This study was performed to develop a rapid immuno-assay kit, by using a specific antigen to detect Hanwoo brand meat. We selected a synthetic antigen specific to our target antibody, named BIO-TAG (Tyr-D-Ala-Phe), by utilizing a computer-based analysis and literature review. BIO-TAG tagged with adjuvant was subcutaneously injected in sheep and Hanwoo. The serum and meat juice of the immunized or non-immunized animal were then analyzed, to measure the titer of antibody by ELISA and Western blot. The amount of antibodies against the BIO-TAG increased (p<0.05) in serum by vaccination. Furthermore, meat juice from the immunized Hanwoo showed greater (p<0.05) antibody titer, compared with those from non-immunized groups. To optimze the dilution factor, we performed dot-ELISA, with various combination levels of BIO-TAG. Results from dot-ELISA showed that 2 mg/mL BIO-TAG was sufficient to distinguish the immunized meat from non-immunized groups. These results support our hypothesis that simple immunization of Hanwoo generates a sufficient amount of antibodies to be detectable in the meat juice by means of the immune-assay. Therefore, specific Hanwoo brand meat can be more precisely identified by our rapid diagnostic kit. This technology can deter possible fraud of counterfeit meat brands in the Korean domestic market with ease and rapidity; and offers a new tool that guarantees consumers high quality Hanwoo brand beef.

• Journal of Plant Biology
• /
• v.37 no.1
• /
• pp.77-83
• /
• 1994

DNA uptake in dry embryos of rice by DNA imbibition was detected by monitoring the expression of chimeric vectors. The selective markers of expression vectors used were ${\beta}-glucuronidase$ ronidase (GUS) and hygromycin phosphotransferase (HPT) genes under the control of CaMV35 S promoter. Frequency of transient expression of the foreign gene was generally 30-50% varying according to the types of vectors and rice cultivars. Dot blot analysis and DNA sequence analysis of inverse polymerase chain reaction products showed that selected rice in hygromycin B (HmB) medium had HPT gene and CaMV35S promoter DNA sequence in genomic DNA of rice. To investigate what ratio of rice having two marker genes simultaneously as rice embryos imbibed the vector DNA having two HPT and GUS gene, transform ants selected in lImB medium were subjected to PCR for GUS gene. It was shown that about 90 percentage of surviving ones in HmB medium had GUS gene.S gene.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.6
• /
• pp.743-749
• /
• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

• International Journal of Oral Biology
• /
• v.35 no.1
• /
• pp.13-19
• /
• 2010

The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.