• Korean Journal of Microbiology
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• v.39 no.2
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• pp.114-117
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• 2003

In molecular biology, it is necessary to develop an easy and rapid method to identify a specific DNA sequence. Though Southern and Northern blot techniques have been used widely for the analysis of gene structure and function, those methods are inconvenient in the points that we need to control incubation temperature, time, and other parameters to get the final result. In this study, we report a new method for the rapid analysis of specific DNA sequence with the modification of an immunochromatographic method. The lateral flow DNA analysis strip is composed of a sample pad, a nitrocellulose membrane for the separation and propagation of analytes, and an absorption pad for the generation of capillary action. Capture DNA was immobilized on the membrane by UV cross-linking and target DNA was labeled with Cy-5 for signaling. The samples containing target DNA were applied onto the sample pad, incubated for 15 min for separation, and scanned with a GSI fluorescence scanner. Though the hybridization reaction occurs in a short time without any washing steps, there appears to be little cross hybridization between the different sequences. The result showed a possibility that the new method can be used for the rapid identification of specific DNA sequence among the samples.

• Korean Journal of Veterinary Pathology
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• v.1 no.1
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• pp.26-32
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• 1997

Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

• 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.

• Molecules and Cells
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• v.26 no.6
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• pp.621-624
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• 2008

We previously showed that Asb-4 and Asb-17 is uniquely expressed in developing male germ cells. A recent report showed that Asb-9 is specifically expressed in the kidney and testes; however, detailed expression patterns in developing germ cells have not been shown. Northern blot analysis in various tissues demonstrated that mAsb-9 was strongly expressed in the testes. Expression analysis by RT-PCR and Northern blot in developing mouse testes indicates that mAsb-9 is expressed from the fourth week after birth to adulthood, with the highest expression in round spermatids. Expression sites were further localized by in situ hybridization in the testes. Pachytene spermatocytes and spermatids expressed mAsb-9 but spermatogonia and generated spermatozoa did not. This study reveals that mAsb-9 could be a specific marker of active spermatogenesis and would be useful for studies of male germ cell development.

• Journal of Plant Biology
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• v.35 no.3
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• pp.253-257
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• 1992

Potyvirus group is the largest group among plant virus groups and damages severely plant hosts upon infectiQn. In order to investigate the mechanism by which potyviruses induce disease in plants, a cDNA clone 29-6 which is cOIlsidered to be a cDNA clone for garlic mosaic virus (GMV) was isolated. It did not hybridize to garlic latent virus genome, which is one of two major garlic viruses. Northern blot analysis shows that the genome size of garlic mosaic virus was about 9 kb. Clone 29-6 strongly hybridizes to poly(A) RNA isolated from garlic leaves, suggesting that GMV RNA is polyadenylated as other potyviruses. Nucleotide sequence analysis of cDNA clones overlapping with clone 29-6 showed that garlic plants are infected with various strains of garlic mosaic virus which are closely related to each other. other.

• Animal cells and systems
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• v.2 no.1
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• pp.113-116
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• 1998

A cDNA clone coding for ribosomal protein 46 (rp46) which is a component of 60S ribosomal large subunit has been identified from Drosophila melanogaster. A cDNA clone encoding S. cerevisiae rp46 was used as a probe to screen a Drosophila larvae cDNA library. The DNA sequence analysis revealed that the cDNA coding for Drosophils rp46 contains a complete reading frame of 153 nucleotides coding for 51 amino acids. The deduced amino acid sequence showed 71-75% homology with those of other eukaryotic organisms. Northern blot analysis showed that about 1-kb rp46 transcripts are abundant throughout fly development. Whole mount embryonic mRNA in situ hybridization also showed no preferential distribution of the transcripts to any specific region. The chromosomal in situ hybridization revealed that the identified gene is localized at position 60C on the right arm of the second polytene chromosome with a possibility of single copy.

• Journal of Microbiology
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• v.33 no.2
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• pp.149-153
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• 1995

Mitochondrial DNA has been isolated from Trimorphomyces papilionaceus. By analyzing DNA fragments digested by restriction enzymes, a restriction site map has been constructured. The mtDNA of T. papilionaceus amounts to 48.5 kb in size and is circular in structure. Entire mitochondrial DNA was cloned in E coli plasmids and Northern blot hybridization was done using cloned and subcloned DNAs as probes. Based on hybridization results of mitochondrial RNA transcripts, a transcription map was prepared.

• Korean Journal of Breeding Science
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• v.41 no.2
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• pp.73-83
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• 2009

To investigate genes related to vernalization and cold- resistance in barley (Hordeum vulgare L. cv. Nagaoka), differentially expressed genes were identified from cold-resistant barley leaves with suppression subtractive hybridization (SSH) and Northern blot analyses. The nucleotide and the deduced amino acid sequences of the putative gene products were compared. The bvrn-7 showed high homology(84%) with gene related to vernalization, and the bvrn-3, bvrn-12, bvrn-28, bvrn-29 and bvrn-36 related to cold-resistant genes had high identity of 88~98% with low temperature-induced genes. The results indicate that the 6 genes were closely related to vernalization and cold-resistance during low temperature treatment.

• The korean journal of orthodontics
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• v.26 no.1 s.54
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• pp.83-94
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• 1996

Substance P is one of the neuropeptide which presents highly in tension site of periodontal ligament during the orthodontic tooth movement. It has bnn also hon as one of the neuropeptides which cause neurogenic inflammation in various tissues and organs. However, there is no report about the effect of substance P on major extracellular matrix protein, collagen production. The purpose of this study was to evaluate the collagen production by substance P in human periodontal ligament cell. The collagenase-digestion method was used to evaluate collagen production and also used Northern blot hybridization for the evaluation of collagen mRNA level. This study also Included in terms of prostanglandins and gelatinase production with respect to collagen production. For the collagen degradation, zymography was used to estimate denatured collagen degradation. Dose-dependent effect of substance P on noncollagen protein, collagen, and percent collagen was that substance P increased noncollagen protein synthesis, but decreased collagen sytnsis. So the percent collagen, which determined by relative collagen production against total protein production, w3s decreased from $7\%\;to\;3.6\%$. This inhibitory effect of substance P on collagen production was disappeared when cells were treated concomitantly with indomethacin. It means that substance P-induced inhibitory effect on collagen production was due at least in part to the production of prostaglandins. To evaluate whether substance P-induced inhibitory effect on collagen production is correspond to the steady-state levels of procollagen mRNA, Northern blot hybridization was performed and it showed that substance P has no effect on the steady-slate level of ${\alpha}1(I)$ procollagen mRNA. It means that the inhibitory effect of substance P on collagen production was due to the change of a certain mechanism after posttranscription. In this context, gelatinase production by substance P in periodontal ligament cells was evaluated by zymography. Zymogram showed that substance P has no effect on gelatinase production in periodontal ligament cells. To explore wheter substance P-induced inhibitory effect on collagen production is selevtive in periodontal ligament cells or not, MC3T3-E1 cells which originated from mouse calvaria was used. It showed that substance P has no effect on collagen production in MC3T3-E1 cells. Taken together, substance P inhibits collagen production in human periodontal ligament cells. This effect was not due to the change of the steady-state level of procollagen mRNA and gelatinase production, but due at least in part to the change of prostaglandins production.

• Journal of Plant Biology
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• v.38 no.2
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• pp.137-141
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• 1995

We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.