• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.139.1-139
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• 2003

Tissues from woody plant contain higher amount of phenolic compounds and polysaccharides, which give inhibitory effects on reverse transcriptase and/or Taq ploymerase. The common multiple-step protocols using several additives to inhibit polyphenoic compounds during nucleic acid extraction are time consuming and laborious. Sodium sulfite (Na$_2$SO$_3$) was used as inhibitor of polyphenolic oxidases in extraction buffer and compare it's effect between commercial RNA extraction kit and small-scale double-stranded RNA (dsRNA) extraction by RT-PCR. During nucleic acid extraction procedure, addition of 0.5％-1.5％ (w/v) sodium sulfite to Iysis buffer or STE buffer resulted in lighter color change than extracts without sodium sulfite and improve the RT-PCR detection. When commercial RNA extraction kit used, optimal concentration of sodium sulfite were variable according to the host plant. However, using dsRNA as RT-PCR template, 1.5％ sodium sulfite in STE buffer improves the detection of both viruses and unspecific amplifications were reduced significantly, Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

• Journal of Life Science
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• v.13 no.1
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• pp.105-109
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• 2003

Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

• International Journal of Oral Biology
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• v.31 no.3
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• pp.73-79
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• 2006

Tissue stem cells are used for the regenerative medicine. In previous study we observed hard tissue formation of human dental pulp-derived cells using alginate scaffold. In this study, we explore the ability to differentiate of the 13th passage cells with glycerol 2-phosphate disodium salt hydrate (${\beta}-GP$) which accelerate calcification. Reverse transcriptase Polymerase Chain Reaction (RT-PCR), transplants using alginate scaffold and histological examination were performed. We observed the expression of DSPP mRNA on day 10 cultured cells with ${\beta}-GP$. In conclusion, the 13th passage cells still have an ability to differentiate into odontoblast-like cells and alginate supports the differentiation of cultured cells in the transplants.

• Advances in pediatric surgery
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• v.5 no.1
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• pp.45-52
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• 1999

Pediatric solid tumors have many histologic similarity. These tumors contained small round cell types, and cause frequent diagnostic problems in pediatric pathology. An important advance in the differentiation of these small round cell tumors has been the identification of consistent chromosomal translocations associated with several types of tumors. Eighteen patients with soft tissue sarcoma were available for review. Seventeen cell lines were also included in this study. The RNA from the specimens were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). PAX3-FKHR fusion was present in four of five alveolar rhabdomyosarcoma and PAX7-FKHR fusion was detected in one of five alveolar rhabdomyosarcoma. None of the specimens expressed more than one chimeric transcript. EWS-FLI1 or EWS-ERG fusions were detected in all seven Ewings' sarcoma. No specimens showed EWS-WT1 fusion. These results corresponded well to the histopathologic diagnosis. There were no differences in the histologic appearances of tumors with the more frequent PAX3-FKHR or EWS-FLI1 fusions compared with those containing the variant PAX7-FKHR or EWS-ERG fusions. RT-PCR assay for chimeric transcript is a useful tool for rapid and objective diagnosis of pediatric solid tumors. Through these tools, we can approach genetically to the differential diagnosis of undifferentiated small round tumors.

• BMB Reports
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• v.44 no.8
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• pp.512-516
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• 2011

Establishment of immortalized human dermal papilla cells (DPCs) retaining the characteristics of DPCs would be a great help for hair researchers. We recently established a simian virus 40T (SV40T)-transformed human DP cell line (SV40TDPC). However, the cell line senesced around passage 25 and ceased proliferation. In this study, we introduced the human telomerase reverse transcriptase (hTERT) gene into SV40T-DPC and established an immortalized human DP cell line. The cell line, SV40T-hTERT-DPC, did not induce tumors when inoculated into nude mice. SV40T-hTERT-DPC maintained morphology of early passage DPCs, expressed markers of DPCs, and retained responses to Wnt/${\beta}$-catenin and bone morphogenic protein (BMP) signaling pathways known to be required for hair-inducing activity of DPCs. The data strongly suggest that SV40T-hTERT-DPC retains many characteristics of human DPCs in vivo without malignant transformation.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.1
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• pp.22-27
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• 2017

The length of hospital stay (LOS) for patients with respiratory virus infections has been reported to depend the virus type and infection severity. However, the impact of co-infections remains unclear. Patients with suspected respiratory virus infections, who visited Dankook University Hospital between December 2006 and February 2014, were included to examine the relationship between co-infections and LOS. Multiplex reverse transcriptase-polymerase chain reactions were used to identify the causative viruses. LOS was analyzed with respect to sex, age, virus, and co-infection. During this period, 5,310 out of the 8,860 patients (59.9%; median age, 1.5 years) were respiratory virus-positive. In respiratory virus-positive patients with single, double, and three-or-more infections, the average LOS was 7.3, 6.7, and 6.6 days, respectively. Longer LOS was observed for older patients and those with human coronavirus OC43 infections compared with adenovirus or respiratory syncytial virus A infections. LOS differed significantly according to age, virus type, and co-infection, but not between double and three or more infections.

• BMB Reports
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• v.38 no.2
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• pp.206-210
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• 2005

Gonadogenesis is a complicated process which involves multi-gene interactions. A rainbow trout (Oncorhynchus mykiss) gene spermatogenesis associated 4 (SPATA4) was cloned and characterized from adult rainbow trout testis. The cDNA sequence of rainbow trout SPATA4 contains an open reading frame of 1, 081 nucleatides encoding a putative protein of 259 amino acids. The putative protein from rainbow trout shares a 76.8% homology with zebrafish SPATA4. No trans-membrane regions or signal peptide were detected using bioinformatics methods. Subcellular localization analysis revealed that rainbow trout SPATA4 was a nuclear protein with highest possibility (39.1%). Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the distribution of rainbow trout SPATA4 in eleven organs of adult rainbow trout. The result demonstrated that this gene express specifically in testis and slight amount of expression was detected in ovary. Further analysis of SPATA4 characterization and function in rainbow trout may provide insight into the understanding of gonadogenesis process.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.167-173
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• 2005

To investigate the possible molecular mechanism (s) of bee venom as a candidate of anti-cancer drug, we examined the effects of the compound on the growth of human lung carcinoma cell line A549. Bee venom treatment declined the cell growth and viability of A549 cells in a concentration-dependent manner, which was associated with induction of apoptotic cell death. Bee venom down-regulated the levels of anti-apoptotic genes such as Bcl-2 and Bcl-XS/L, however, the levels of Bax, a pro-apoptotic gene, were up-regulated. Bee venom treatment induced not only tumor suppressor p53 but also cyclin-dependent kinase inhibitor p21WAF1/CIP1 expression in a dose-dependent manner. Furthermore, bee venom treatment induced the down-regulation of telomerase reverse transcriptase mRNA and telomeric repeat binding factor expression of A549 cells, however, the levels of telomerase-associated protein-1 and c-myc were not affected. Taken together, these findings suggest that bee venom-induced inhibition of human lung cancer cell growth is associated with the induction of apoptotic cell death via regulation of several major growth regulatory gene products, and bee venom may have therapeutic potential in human lung cancer.

• The Plant Pathology Journal
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• v.30 no.4
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• pp.445-449
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• 2014

The incidence of Cherry necrotic rusty mottle virus (CNRMV) and Cherry green ring mottle virus (CGRMV) have recently been occurred in Korea, posing a problem for sweet cherry cultivation. Since infected trees have symptomless leaves or ring-like spots on the pericarp, it is difficult to identify a viral infection. In this study, the incidence of CNRMV and CGRMV in sweet cherry in Gyeongbuk province was surveyed using a newly developed duplex reverse transcriptase polymerase chain reaction (RT-PCR) method that can detect both viruses in a single reaction. CNRMV and CGRMV co-infection rates were 29.6%, 53.6%, and 17.6%, respectively, in samples collected from three different sites (Daegu, Gyeongju and Gyeongsan) in Gyeongbuk province during 2012 and 2013. This duplex RT-PCR method offers a simple, rapid, and effective way of identifying CNRMV and CGRMV simultaneously in sweet cherry trees, which can aid in the management of viral infections that could undermine yield.

• Korean Journal of Veterinary Research
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• v.41 no.3
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• pp.333-342
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• 2001

To detect the genetic variations among infectious bursal disease (IBD) vaccine strains, the hypervariable region of VP2 gene of seven IBDV vaccine strains were amplified using reverse transcriptase/polymerase chain reation(RT/PCR). Ampllified PCR products of IBDV were cloned, sequenced, and compared with published sequences for IBDV. Vaccine strains (JOONG, HAN, B7, IB, BU2, G2, CIL) used in Korea and Korean field isolates (SH/92, K1, 310) had 81%(310 and HAN) ~ 98%(SH/92 and CIL) amino acid sequence similarity. Vaccine strains had 80%(HAN and IB) ~ 99%(JOONG and BU2) amino acid sequence similartiy. Intermediate plus vaccine strain, CIL was not substituted at positions 279(D $\rightarrow$ N) and 284(A $\rightarrow$ T), and conserved in serine-rich heptapeptide. At the two hydrophilic region, JOONG, IB and Bu2 strains had identical amino acid sequence comparing with STC strain. By phylogenetic analysis, JOONG and DAE strains were categorized in same group with BU2. The CIL and STC strains closely related but seperated from G2, HAN, B7 and IB strains.