• Journal of Gastric Cancer
• /
• v.1 no.4
• /
• pp.202-209
• /
• 2001

Purpose: When cancer cels invade the stroma, they should be dissociated from the adjacent cells at first. E-cadherin and $\beta-catenin$ constitute an important protein complex associated with cellular adhesion, development, and differentiation, especially in epithelial cells. The role of E-cadherin and $\beta-catenin$ in gastric carcinogenesis were studied. Materials and Methods: The expression of E-cadherin and $\beta-catenin$ in gastric adenocarcinomas by using immunohistochemical staining and the mutation by using polymerase chain reaction- single stranded conformation polymorphism (PCR-SSCP) and sequencing were performed in 40 adenocarcinomas and 5 dysplasia of stomach. Thirteen cases, which had lymph node metastasis, were also included for immunohistochemical staining. Results: Inappropriate cytoplasmic and/or nuclear expression of a E-cadherin-$\beta-catenin$ complex was more frequent in poorly differentiated, diffuse type signet ring cell carcinomas than in well-differentiated, intestinal type adenocarcinomas (P<0.05). However, the expression was not related with clinical stage or lymph node metastasis. Mutation of E-cadherin was detected in 4 cases by using PCR-SSCP, whereas mutation of $\beta-catenin$ was detected in 2 cases. Conclusion: E-cadherin and $\beta-catenin$ seem to be important in gastric carcinogenesis, especially in poorly differentiated diffuse type.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.1
• /
• pp.7-12
• /
• 2010

The full-length cDNA of the porcine peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase (PECI) gene encodes a monofunctional peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase. Cloning and sequencing of the porcine PECI cDNA revealed the presence of an 1185-base pair open reading frame predicted to encode a 394-amino acid protein by the 5'rapid amplification of cDNA ends (5'RACE) and EST sequences. The porcine PECI gene was expressed in seven tissues (heart, liver, spleen, lung, kidney, skeletal muscle, fat) which was revealed by reverse transcriptase-polymerase chain reaction (RT-PCR). The porcine PECI was mapped to SSC71/2 p11-13 using the somatic cell hybrid panel (SCHP) and the radiation hybrid panel (RH) (LOD score 12.84). The data showed that PECI was closely linked to marker S0383. A C/T single nucleotide polymorphism in PECI exon 10 (3'UTR) was detected as a PvuII PCR-RFLP. Association analysis in our experimental pig population showed that different genotypes of PECI gene were significantly associated with the Average Backfat thickness (ABF) (p<0.05) and Buttock backfat thickness (p<0.01).

• KSBB Journal
• /
• v.25 no.3
• /
• pp.215-222
• /
• 2010

Monogenic diseases are resulted from modifications in a single gene of human cells. Because their treatment with pharmacological medicine have a temporary effect, continuous nursing care and retreatment are required. Gene therapy, gene targeting and induced pluripotent stem cell (iPSC) are considered permanent treatment methods of them. In gene therapy, however, retroviral vectors that have potential toxicity caused by random insertion of harmful virus are used as vehicles for transferring genetic materials. On the other hand, gene targeting could replace and remove the modified gene though homologous recombination (HR) induced by site-specific endonucleases. This short review provides a brief overview on the recently tailored endonucleses with high selectivity for HR.

• Journal of the Korean Society of Fisheries and Ocean Technology
• /
• v.50 no.4
• /
• pp.542-550
• /
• 2014

We investigated biomass, diatom species and fucoxanthin contents as cell growth, fatty acid and amino acid contents as nutritional composition of diatoms attached on plate to confirm effects of light emitting diodes (LEDs) due to block off natural light. In the single LED irradiation, biomass showed significantly higher to $30.0{\pm}6.48mg/m^2$ in white LED than that of others (P<0.05). The dominate diatom species was Navicula cancellata. Their lipid contents showed significantly higher to $112.9{\pm}19.23ug/mg$ dry matter (DM) in control than that of others LEDs. But eicosapetaenoic acid (EPA) contents showed significantly higher to $3.3{\pm}0.62ug/mg$ DM than others, but not significantly differed with natural control light treatment (P<0.05). And total protein contents are higher in control and blue LED light than that of others, but essential amino acid contents showed significantly higher to $3.2{\pm}4.8%$ in control (P<0.05). In mixing light with natural and LED light, biomass showed $2.6{\pm}0.22mg/m^2$ in blue LED (P<0.05). Fatty acids contents were not significantly differed with all treatments. Amino acid contents showed to $11.0{\pm}0.33ug/mg$ DM in white LED (P<0.05), but not significantly differed with others LED lights (P>0.05). Therefore, we could suggest that irradiation of blue LED in natural light very benefit to diatom culture for larvae of sea cucumber and abalone and do on.

• Microbiology and Biotechnology Letters
• /
• v.2 no.1
• /
• pp.1-7
• /
• 1974

For the purpose of producing cellulosic single-cell protein from the agricultural wastes, 172 strains of cellulose-assimilating bacteria were isolated from 102 samples of rotten woods, compost soils, soils and so on by the enrichment culture technique. The isolates were examined for their ability to utilize cellulose as carbon source, and then six strains were screened by their strong cellulose assimilating ability and identified as follows: 1. Among six strains of bacteria screened, five strains were identified as species belonged to the genus Cellulomonas and the remainder to the genus of Sporocytophaga. 2. The isolated Sporocytophaga species was identified as S. ellipsosporn because it has a ellipsoidal microcyst. 3. The isolated Cellulomonas species were identical to a strain of C. fimi, C. aurcgena, C. gelida, respectively and two strains to C. flavigena. 4. The isolated C. aurogena was proved to be a new variety becauuse it has different characteristics of assimilating pentoses such as arabinose and xylose from the strain discribed in Bergey's Manual.

• BMB Reports
• /
• v.33 no.4
• /
• pp.337-343
• /
• 2000

The human immunodeficiency virus type 1 (HIV-1), transactivator of transcription (Tat), is one of the viral gene products that is essential for HIV-1 replication. The HIV-l Tat protein regulates transcription from an HIV-1 long terminal repeat (LTR) and affects the gene expression of cellular proteins during infection. In order to develop an expression system to overexpress and simply purify HIV-1 Tat proteins, the HIV-1 Tat coding sequences that contain one or two exons were amplified using PCR and cloned into a pET vector, which contains a consecutive stretch of six histidine residues at the amino-terminus. The reconstituted vectors were overexpressed in the E. coli strain and the soluble recombinant proteins were purified to be homogeneity in a single step by $Ni^{+2}-nitrilotriacetic$ acid Sepharose chromatography under nondenaturing conditions. Recombinant HIV-1 Tat proteins were shown to transactivate the HIV-1 LTR promoter in a dose-dependent manner when introduced into mammalian cells. In addition, treatment of human endothelial cells with purified Tat proteins resulted in a significant increase in the level of vascular cell adhesion molecule-1 (VCAM-1) expression. These results indicate that the recombinant HIV-1 Tat proteins are active in transactivating viral and cellular promoters. The expression and purification system described in this study will facilitate in characterizing the biological functions of the Tat proteins.

• The Korean Journal of Physiology and Pharmacology
• /
• v.24 no.3
• /
• pp.277-286
• /
• 2020

Polycystic kidney disease 2-like-1 (PKD2L1), also known as polycystin-L or TRPP3, is a non-selective cation channel that regulates intracellular calcium concentration. Calmodulin (CaM) is a calcium binding protein, consisting of N-lobe and C-lobe with two calcium binding EF-hands in each lobe. In previous study, we confirmed that CaM is associated with desensitization of PKD2L1 and that CaM N-lobe and PKD2L1 EF-hand specifically are involved. However, the CaM-binding domain (CaMBD) and its inhibitory mechanism of PKD2L1 have not been identified. In order to identify CaM-binding anchor residue of PKD2L1, single mutants of putative CaMBD and EF-hand deletion mutants were generated. The current changes of the mutants were recorded with whole-cell patch clamp. The calmidazolium (CMZ), a calmodulin inhibitor, was used under different concentrations of intracellular. Among the mutants that showed similar or higher basal currents with that of the PKD2L1 wild type, L593A showed little change in current induced by CMZ. Co-expression of L593A with CaM attenuated the inhibitory effect of PKD2L1 by CaM. In the previous study it was inferred that CaM C-lobe inhibits channels by binding to PKD2L1 at 16 nM calcium concentration and CaM N-lobe at 100 nM. Based on the results at 16 nM calcium concentration condition, this study suggests that CaM C-lobe binds to Leu-593, which can be a CaM C-lobe anchor residue, to regulate channel activity. Taken together, our results provide a model for the regulation of PKD2L1 channel activity by CaM.

• The Journal of the Korea Contents Association
• /
• v.10 no.8
• /
• pp.104-110
• /
• 2010

In the field of biotechnology, computer can play varied roles such as the ordinal analysis, ordianl comparison, nutation tracing, analogy comparison for drug design, estimation of protein function, cell mechanism, and verifying the role of a gene for preventing diseases. Additionally, by constructing database, it can provide an application for the cloning process in other data researches, and be used as a basis for the comparative genetics. For the most of researcher about biotechnology, they need to use the tool that can do all of job above. This study is focused on looking into problems of existing systems to analysis bio data, and designing an improved analyzing system that can propose a solution. In additional, it has been considered to improve the performance of each constituent, and all the constituents, which have been separately processed, are combind in a single system to get over old problems of the existing system.

• The Korean Journal of Physiology and Pharmacology
• /
• v.9 no.2
• /
• pp.131-135
• /
• 2005

Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.9
• /
• pp.1237-1241
• /
• 2005

Mannose-P-dolichol utilization defect 1 (MPDU1) gene is required for utilization of the mannose donor MPD in synthesis of both lipid-linked oligosaccharides (LLOs) and glycosylphosphatidylinositols (GPI) which are important for functions such as protein folding and membrane anchoring. The full length cDNA of the porcine MPDU1 was determined by in silico cloning and rapid amplification of cDNA ends (RACE). The deduced amino acid showed 91% identity to the corresponding human sequence with five predicted transmembrane regions. RT-PCR was performed to detect its expression pattern in five tissues and results showed that it is expressed ubiquitously among the tissues checked. A single nucleotide substitution resulting in the amino acid change (137 Tyr-137 His) was detected within exon 5. Allele frequencies in six pig breeds showed distinctive differences between those Chinese indigenous pigs breeds and European pigs. Using the pig/rodent somatic cell hybrid panel (SCHP), we mapped the porcine MPDU1 gene to SSC12, which is consistent with the comparative mapping result as conservative syntenic groups presented between human chromosome 17 and pig chromosome 12.