• Journal of Microbiology
• /
• v.38 no.4
• /
• pp.230-238
• /
• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.1 no.1
• /
• pp.29-33
• /
• 2000

We cloned and characterized a very late expression factor 1 gene, vlf-1, which regulates the level of very late gene transcripts, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 1,140 bp vlf-1 has an open reading frame of 379 amino acid and a MW of 44 kDa. The vlf-1 nucleotide sequence of BmNPV-Kl showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain so far known, and its deduced amino acid residues were identical to those of BmNPV T3. The location of vlf-1 in the BmNPV-Kl genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level were confirmed by Northern hybridization analysis.

• Journal of Plant Biology
• /
• v.34 no.2
• /
• pp.121-127
• /
• 1991

Total soluble proteins from three developmental stages of induced root nodules of Canavalia lineata were compared with those of non-nodulated roots by SDS-PAGE and two dimensional (2-D) gel electrophoresis. Thirteen nodule-specific protein (nodulin) bands were identified by the former and 30 nodule specific protein spots were detected by the latter method respectively. Some of the nodulins were detected differentially depending on the nodule's developmental stages. For example, only three leghemoglobin (Lb)-like protein spots appeared at stage I (d<2 mm), but two additional Lb-like protein spots appeared at stage II (d <4-5 mm). pI value and molecular weight of nomomers of Lb-like protein were narrower and greater than those of soybean, ranging from 4.4 to 5.0 and 15.7 kd respectively. Northern blot hybridization of total RNAs from roots and root nodules using soybean Lb cDNA as a probe made it clear that Lb gene was expressed tissue-specifically only in the root nodules.odules.

• Applied Biological Chemistry
• /
• v.37 no.4
• /
• pp.248-254
• /
• 1994

Bromoxynil is an antidicot herbicide widely used on cereal crops and has a short half life in the soil. A bxn gene, encoding a specific nitrilase that converts bromoxynil to its primary metabolite 3,5-dibromo-4-hydroxybenzoic acid, was inserted in plant binary vector pGA482, and then introduced into tobacco and lettuce plants via Agrobacterium mediated leaf-disc transformation method. Transgenic plants with the bxn gene were selected by kanamycin and regenerated to whole plants. The regenerated transgenic plants were determined level of expression of bxn gene by Northern blot analysis. Leaf-disc analysis and pot-assay confirmed that the transgenic tobacco and lettuce plants were resistant to high doses of bromoxynil.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.5
• /
• pp.1152-1157
• /
• 2005

The effect of carbon sources on nisin Z biosynthesis in Lactococcus lactis subsp. lactis A164 was studied in batch culture using M17 broth containing different carbon sources. Among the eleven carbon sources tested, glucose, sucrose, and lactose were suitable carbon sources for cell growth of L. lactis A164. In particular, cells grown on lactose produced at least 3-fold greater amount of nisin Z than those on other carbon sources. Galactose resulted in less amount of cell mass than did sucrose or glucose, but gave a higher level of nisin Z activity. Northern blot analysis revealed. that lactose increased the transcription of the nisZ pre-peptide gene. Although galactose was less efficient than lactose, it increased the transcription of nisZ along with a higher level of nisin Z than did sucrose and glucose. These results suggest that the increased nisin Z production is correlated with the induction of nisZ by lactose and galactose. Among all the carbon sources tested, no remarkable differences were observed in nisRK and nisFEG transcripts, indicating that the lactose- or galactose-mediated induction is unique to the nisZ promoter.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.28 no.6
• /
• pp.434-439
• /
• 2002

Oral squamous cell carcinoma (OSCC) is one of the most common head and neck cancers. OSCC generally has a poor prognosis due to its tendency towards a local invasion and subsequent metastasis, which is mediated by multiple proteolytic enzymes and angiogenesis. Soy products contain high levels of isoflavonoids, including the tyrosine kinase inhibitor, genistein, which has been identified as a potent inhibitor of cell proliferation and in vitro angiogenesis. The purpose of this in vitro study is to evaluate the anti-cancer effect of genistein with respect to the angiogenesis and basement membrane invasion in OSCC. The highly invasive OSCC cell line, HSC-3 cells were cultured in the presence of $10{\mu}M$ genistein for 24h. To evaluate the effects of genistein on the invasiveness and the gelatinolytic activity, in vitro invasion assay and zymography were performed. In order to evaluate the effect on the VEGF and bFGF mRNA expression, RT-PCR and northern hybridization reaction, and chemiluminescence detection were applied. The in vitro invasion assay showed that the genistein treatment reduced the cellular invasion through the artificial basement membrane and significant difference between the control group and the genistein treated group was shown in MMP-2 activity. Especially, the 62 kDa activated form of MMP-2 in the control group was 1.8 times higher than that in the genistein treated group. The results of the northern blot analyses indicated that VEGF mRNA expression in the genistein treated group was significantly down regulated. This study showed that genistein inhibits angiogenesis and reduces basement membrane invasion in OSCC. It seems to support the possibility of genistein as an anti-cancer agent.

• Journal of Sericultural and Entomological Science
• /
• v.52 no.1
• /
• pp.39-44
• /
• 2014

We isolated highly-expressed genes in the posterior silk glands of silkworm on a previously study, which one of these was identified as RNA binding protein-1 homologue (RBP-1) gene. In this study, we investigated gene expressional characteristics of the RBP-1 depending on silkworm development stages and several tissues of the larvae, respectively. Northern blot hybridization analysis showed that the RBP-1 gene was expressed high in larval and pupal periods, and highly expressed than endogenous internal control gene (BmA3) on all tested larval tissues. In addition, we isolated and analyzed a phage DNA having 1,660 bp-long promoter region of the RBP-1 gene from a genomic DNA library. To study the RBP-1 gene promoter activity, RBP-1 (-740/+ 30) was amplified by PCR and subcloned into a pGL3 basic vector to generate pGL-RBP1. A luciferase report vector carrying RBP-1 gene promoter (770 bp) was tested by luciferase assay in Sf9 cells. In the result, the RBP-1 gene promoter was more efficient than constitutive promoter (BmA3) by approximately ten percent.

• Animal Systematics, Evolution and Diversity
• /
• v.15 no.2
• /
• pp.153-164
• /
• 1999

One hundred and eleven samples of six subspecies of striped field mouse, Apodemus agrarius Pallas from Korea, China and Russia, were used for the analysis of mitochondrial DNA(mtDNA) fragment patterns resulted from the digestion with eight restriction enzymes by blot hybridization technique. All 32 fragments, nine mtDNA haplotypes, and four major subgroups with the mean divergence value of 0.896 to 1.150% were revealed. In summary, three forms are recognized: [I, subspecies chejuensis (Chejudo island, Korea)], [II, subspecies pallescens (southwestern Korea), coreae (central Korea), and septentrionalis (Russia)], and [III, subspecies manchuricus (northeastern China) and pallidior (northern China)], although some samples of subspecies coreae are somewhat different from almost all samples of six subspecies, and some samples of subspecies pallidior are similar with all samples of subspecies septentrionalis to form same haplotype. It is confirmed that A. agrarius chejuensis is a distinct subspecies, that subspecies coreae (including pallescens) is also a distinct subspecies, that subspecies manchuricus and pallidior are synonyms of subspecies ningpoensis, and that subspecies septentrionalis is a synonym of subspecies ningpoensis, and that subspecies septentrionalis is a synonym of subspecies agrarius. Moreover, it seems that A. agrarius shows constant karyotype, minimal variation in mtDNA genotype, and considerable divergence in morphometric characters, although further analyses with additional samples of A. agrarius in Eurasia will be necessary to determine the degree of variation of these taxonomic characters and to clarify subspecies classification as well.

• Journal of Aquaculture
• /
• v.19 no.3
• /
• pp.157-165
• /
• 2006

Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.

• Journal of Life Science
• /
• v.14 no.6 s.67
• /
• pp.1023-1027
• /
• 2004

The RAD3 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. An yeast RAD3 gene has been previously isolated by functional complementation. In order to identify the RAD3 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD3 DNA, and then isolated RAD3 homologous DNA from C. cinereus chromosome. The RAD3 homolog DNA was contained in 3.2 kb DNA fragment. Here, we report the results of characterization of a fungus C. cinereus homolog to the yeast RAD3 gene. Southern blot analysis confirmed that the C. cinereus chromosome contains the RAD3 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from the C. cinereus cells were hybridized with the 3.4 kb PvuII DNA fragment of the S. cerevisiae RAD3 gene, transcripts size of 2.8 kb were detected. In order to investigate whether the increase of the amount of transcripts by DNA damaging agent, transcript levels were examined after treating agents to the cells. The level of transcripts were not increased by untraviolet light (UV). This result indicated that the RAD3 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the HRD3 gene is essential for viability of the cells and DNA repair function. These observations suggest an evolutionary conservation of other protein components with which HRD3 interacts in mediating its DNA repair and viability functions.