• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.285-292
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• 1993

C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.203-210
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• 1997

The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

• Tuberculosis and Respiratory Diseases
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• v.70 no.4
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• pp.315-322
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• 2011

Background: A variety of diagnostic modalities for lung cancer have been developed. To achieve efficient and early detection of lung cancer, we tried to measure the expression rates of the melanoma associated gene (MAGE) and synovial sarcoma on X chromosome (SSX) genes. Methods: We designed primers for the SSX gene. In addition to the pre-developed MAGE A primer, using an SSX gene primer was attempted to increase the detection rate. We obtained cancer tissues and cancer-free lung tissues from resected lung, sputum from lung cancer patients who had not undergone surgery, and sputum from healthy people and patients with benign intrathoracic diseases. Results: The sensitivity of the MAGE or SSX gene RT-PCR to identifying cancer tissue of the 69 lung cancer patients was 95.2% for squamous cell carcinoma (scc), 87.0% for adenocarcinoma, and 100% for small cell carcinoma. The mean sensitivity value was 94.2% (p=0.001). For adenocarcinoma, the additional use of the SSX gene resulted in a higher expression rate than MAGE alone (87% vs. 69.6%). The expression rate for the cancer-free lung tissue was 14.3% in scc, 17.4% in adenocarcinoma, and 25.0% in small cell carcinoma. In the induced sputum of 49 lung cancer patients who had not undergone surgery, the expression rate for one of the two genes was 65.5%. The expression rate for the sputum of healthy people and benign intrathoracic diseases by MAGE or SSX gene reverse transcription polymerase chain reaction (RT-PCR) was 3.8% and 17.7%. Conclusion: Detecting lung cancer using the expression of MAGE and SSX genes in lung cancer tissue has high sensitivity.

• Korean Journal of Oriental Medicine
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• v.8 no.1
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• pp.109-126
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• 2002

The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwangtang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by hehavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed $({\sim}100%)$, whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidy lethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK 108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

• Biomedical Science Letters
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• v.10 no.4
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• pp.333-339
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• 2004

A multiplex PCR assay targeting the yst and 16S rRNA genes of Yersinia enterocolitica was developed to specifically identify pathogenic Y. enterocolitica from pure culture. Simultaneous amplification of 145 and 416 bp fragments of the yst and 16S rRNA genes of Y. enterocolitica was obtained using the primer pairs in a single reaction. Validation of the assay was performed with the reference Yersinia strains and other members of the family Enterobacteriaceae. The defined primer pairs amplified the targeted sequence from only pathogenic Y. enterocolitica strains, whereas none of the other bacterial species yielded any amplified fragments. Within an assay time of 4 h, this assay offers a very specific, reliable, and inexpensive alternative to the conventional phenotypic assays used in clinical laboratories to identify pathogenic Y. enterocolitica.

• Journal of fish pathology
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• v.34 no.1
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• pp.31-38
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• 2021

The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H₂S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

• Korean Journal of Oriental Medicine
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• v.9 no.1
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• pp.113-122
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• 2003

In the present paper, morphological characteristic and PCR analysis were carried not to identify the medicinal origins of Peucedanum decursivum and Anthriscus sylvestris. Some samples in Korean. markets were Peucedanum decursivum misused with rhizomes of Anthriscus sylvestris. The main. origin of Peucedanum decursivum producted was quite different from those of Anthriscus sylvestris in the morphological characteristics. And PCR analysis, the three primer has developed into anew tool for identification of Peucedanum decursivum and Anthriscus sylvestris.

• Korean Journal of Poultry Science
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• v.35 no.1
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• pp.9-13
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• 2008

Salmonella enterica serotype gallinarum biovar Gallinarum or Pullorum and Salmonella enterica serotype Enteritidis are the most important diseases in poultry industry. Transitional diagnosis methods of these diseases such as direct isolation and identification by a biochemical test are time consuming with low specificity. In this study, we have focused on the suitable procedure for the rapid and accurate diagnosis of diseases derived from the three Salmonella strains. We initially confirmed Salmonella species by PCR using a specific ITSF/ITSR primer pair instead of biochemical test, and then the PCR-amplified phase 1 flagellin (fliC) using a specific fliCF/fliCR primer pair was digested with a restriction endonuclease, Bpm I and/or Bfa I, to discriminate among S. Pullorum, S. Gallinarum, and S. Enteritidis. We found that these methods could be applied to field isolates of the three Salmonella strains to detect and to discriminate rapidly for convenient diagnosis.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.213-221
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• 2005

We used nine decamer primers to generate DNA fragment sizes ranging from 100 bp to 1,600 bp from two bullhead (Pseudobagrus fulvidraco) populations of Dangjin in Korea. 376 fragments were identified in the cultured bullhead population, and 454 in the population of wild bullhead from Dangjin: 287 specific fragments $(76.3\%)$ in the cultured bullhead population and 207 $(45.6\%)$ in the wild bullhead population. On average, a decamer primer was used to generate 34.2 amplified products in a cultured bullhead. A RAPD primer was used to generate an average of 3.1 amplified bands per sample, ranging between 2.5 and 6.0 fragments in this population. Nine primers also generated 24 polymorphic fragments (24/376 fragment, $6.4\%$) in the cultured bullhead population, and 24 (24/454 fragments, $5.2\%$) in the wild bullhead population. The OPA-16 primer, notably, produced which 11 out of 11 bands $(100\%)$ were monomorphic in the wild bullhead population. 110 intra-population-specific fragments, with an average of 12.2 per primer, were observed in the cultured bullhead population. 99 fragments, with an average of 11.0 per primer, were identified in the wild bullhead. Especially, 55 inter-population-common fragments, with an average of 6.1 per primer, were observed in the two bullhead populations. The bandsharing value (BS value) of individuals within the wild bullhead population was substantially higher than was determined in the cultured bullhead population. The average bandsharing value was $0.596\pm0.010$ within the cultured bullhead population,. and $0.657\pm0.010$ within the wild bullhead population. The dendrogram obtained with the nine primers indicates two genetic clusters, designated cluster $1\;(CULTURED\;01\~CULTURED\;11)$, and cluster $2\;(WILD\;12\~WILD\;22)$. Ultimately, the longest genetic distance displaying significant molecular differences was determined to exist between individuals in the two bullhead populations, namely between individuals WILD no. 19 of the wild bullhead population and CULTURED no. 03 of the cultured bullhead population (genetic distance = 0.714). RAPD-PCR allowed us to detect the existence of population discrimination and genetic variation in Korean population of bullhead. This finding indicates that this method constitutes a suitable tool for DNA comparison, both within and between individuals, populations, species, and genera.

• Korean Journal of Medicinal Crop Science
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• v.9 no.3
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• pp.205-210
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• 2001

Dried parts of the two species are difficult to distinguish morphologically, thus Codonopsis radix has been sold instead of Adenophorae radix in herbal medicine market. Therefore, this study was conducted to develop the genetic marker through the examination of the phylogenetic relationships between two Adenophora triphylla(Thunb.) A. DC. var. japonica Hara, two Adenophora radiatifolia Nakai, five Codonopsis lanceolata(Sieb. et Zucc)Trautv. using RAPD analysis. Fifty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Adenophorae radix and Codonopsis radix. Based on the RAPD patterns, the genetic relationships between three herbal medicine were analyzed by UPGMA method. As a result, Adenophorae radix and Codonopsis radix were classified into two major subgroups on the basis of the genetic similarity coefficient. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. Furthermore, the specific RAPD patterns were produced from the mixture of dried roots of A. triphylla and C. lanceolata. These results prone the usefulness of the RAPD analysis for the discrimination of pure materials from the mixtures of A. triphylla and C. lanceolata.