• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.23-28
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• 2010

Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.71.2-71
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• 2003

Ralstonia solancearum infects many solanaceous plants, however race 3 infects only potato and tomato weakly. To identify genes responsible for race specificity of R. solanacearum, we mobilized genomic library of LSD2029 (race 3) into LSD341 (race 1) and inoculated 1,000 transconjugants into hot pepper. One transconjugant that did not induce wilt symptom in hot pepper was isolated. We found that a cosmid clone, pRSl, conferred avirulence to LSD341. By deletion and mutational analyses of pRSl, we found the 0.9-kb PstI/Hindlll fragment carries avirulence functions. We sequenced the fragment and identified one possible open reading frame, a rsal gene, possibly encoding 110 amino acids. The rsal was preceded with a plant-inducible promoter (PIP) box, indicating that the gene might be regulated by HrpB. Interestingly, the promoter region of the rsal homolog in the strain GM11000 (race 1) did not have the PIP box. Rsal did not show any significant homologies with proteins in the database, indicating th e protein is different from the previously reported avirulence proteins. When we mutated the rsal gene by marker-exchange in LSD2029, the mutant was less virulent in potato.

• The Journal of the Korean Society for Microbiology
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• v.35 no.1
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• pp.49-60
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• 2000

ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.187-193
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• 1996

The T sergenti DNA fragments used as probes of KTS1(2.4kb) and KTS3(1.5kb) were labeled with digoxigenin-11-dUTP for the Southern hybridization. T sergenti DNAs from different geographic locations(Korea; Chonbuk, Kyungbuk, Chungnam, Kangwon, Cheju island, Japan; Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9202, Shintoku 9102) which had been digested with Pst I and EcoR I were probed by the digoxigenin-11-dUTP-labeled KTS1 and KTS3. As the results, the samples from Chonbuk, Kyungbuk, Cheju island in Korea and Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9102 in Japan were positively reacted, but the others from the other locations not reacted. In the comformation test of T sergenti DNA from different geographic locations, all of the samples were positively detected by PCR amplification.

• Journal of Oral Medicine and Pain
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• v.11 no.1
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• pp.67-78
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• 1986

This study was attempted to identify the emotional characteristics of temporomandibular disorder patients. The author applied one of the self-report modes of psychological measurement, Symptom Chechlist-90-Revision. The subjects were 219 TM disorder patients who visited the Department of Oral Diagnosis and Oral Medicine, Seoul National University Hospital during the period from December 1985 to September 1986. All the patients were divided into subgroup according sex, age, duration of symptoms, presence or absence of T-scores of each symptom dimension and global index. The obtained result were as follows : 1. Mean value of T-scores of each symptom dimension and global of the overall patients was within normal range. The two higher mean values of T-scores among 9 symptom dimensions were those of SOM and ANX. 2. Mean values of T-scores of females were higher than those of males in the O-C, DEP, ANX, HOS, PSY dimensions and all global indices, and there was a significant difference in the distribution of T-scores of the SOM dimension between males and female(P<0.05). 3. There was no significant difference between the subgroup under 30 years and the subgroup 30 years or older. 4. The subgroup with symptoms for 6 months or longer showed the higher mean values of T-scorers in the SOM, O-C, I-S, DEP, ANX, PHOB, PAR, PSY dimensions and all global indices compared with the subgroup with symptoms for shorter than 6 months. 5. The subgroup with pain showed the higher mean values of T-scores in all the symptom dimensions except the PAR in comparison with the subgroup with other complaints than pain, and there was a significant difference in the distribution of T-scores of the PST index between the pain subgroup and the non-pain subgroup(P<0.05). There was a significant difference in the distribution of T-scores of the PHOB dimension between the high-school graduates subgroup and the college graduates subgroup(P<0.05).

• Journal of Microbiology
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• v.43 no.spc1
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• pp.110-117
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• 2005

Salmonella enterica serovar Typhimurium causes human gastroenteritis and a systemic typhoid-like infection in mice. A critical virulence determinant of Salmonella is the ability to invade mammalian cells. The expression of genes required for invasion is tightly regulated by environmental conditions and a variety of regulatory genes. The hilA regulator encodes an OmpR/ToxR family transcriptional regulator that activates the expression of invasion genes in response to both environmental and genetic regulatory factors. Work from several laboratories has highlighted that regulation of hilA expression is a key point for controlling expression of the invasive phenotype. A number of positive regulators of hilA expression have been identified including csrAB, sirA/barA, pstS, hilC/sirC/sprA, fis, and hilD. HilD, an AraC/XylS type transcriptional regulator, is of particular importance as a mutation in hilD results in a 14-fold decrease in chromosomal hilA::Tn5lacZY-080 expression and a 53-fold decrease in invasion of HEp-2 cells. It is believed that HilD directly regulates hilA expression as it has been shown to bind to hilA promoter sequences. In addition, our research group, and others, have identified genes (hilE, hha, pag, and lon) that negatively affect hilA transcription. HilE appears to be an important Salmonella-specific regulator that plays a critical role in inactivating hilA expression. Recent work in our lab has been directed at understanding how environmental signals that affect hilA expression may be processed through a hilE pathway to modulate expression of hilA and the invasive phenotype. The current understanding of this complex regulatory system is reviewed.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.1
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• pp.129-141
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• 2001

When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

• The Korean Journal of Food And Nutrition
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• v.8 no.4
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• pp.307-317
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• 1995

The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.138-146
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• 1985

A Bacillus licheniformis ATCC31667 gene coding for a glucose isomerase has been cloned and expressed in glucose isomerase negative mutant of Escherichia coli. A recombinant plasmid, constructed by ligation of a EcoRI fragment of B.licheniformis chromosomal DNA to vector plasmid pBR322, was expressed glucose isomerase positive in E.coli LE392-6 with growth on minimal medium containing xylose as a sole carbon source. This recombinant plasmid, designated pBGI6, had the insery of 4.1Kb of Bacillus gene in EcoRI site, and restriction map of the plasmid was established. The plasmid pBG16 was very stable after 10days of serial transfer to a fresh medium. The activity of glucose isomerase from the transformed cell containing pBGI6 was increased about 20 fold than its wild type of host.

• Journal of Aquaculture
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• v.15 no.1
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• pp.7-13
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• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.