• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.5
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• pp.1075-1082
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• 2014

Monthly Screening of Pacific oyster, Crassostrea gigas, was surveyed by PCR for Ostreid herpesvirus(OsHV-1) infections from 2012 to 2013 in Korea. It was showed positive results in one hatchery in April and May 2012 and in oyster farming areas from August to November in 2013. The obtained sequences showed 99.3~99.5% homology with OsHV-1 ${\mu}var$(Accession Number: HQ842610) and 96.2~96.3% homology with OsHV isolated from Korea(Accession Number: AY509253).

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.8
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• pp.1071-1075
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• 2002

In order to identify and develop the specific DNA marker for the identification of Hanwoo (Korean Cattle) from other breeds, a specific DNA marker of 519 bp was identified and sequenced from polymorphic analysis using RAPD-PCR for 6 cattle breeds. Two different repetitive sequences, $(AAC)_5$ and $(GAAGA)_2$, were selected and designed to use specific probe to develop a DNA marker for Hanwoo specific. When the $(AAC)_5$ probe was applied, the 10 kb specific DNA marker showed in the DNA fingerprinting from 237 of 281 Hanwoo individuals. This novel Hanwoo specific DNA probe is useful to perform the marker-assisted selection for screening Hanwoo purity as an unique genetic source.

• Journal of the Korean Society of Tobacco Science
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• v.25 no.2
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• pp.87-94
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• 2003

Recently, many researches for plant alkaloids, one of the largest groups of natural products, are reported because of their various pharmacological activity. This study was carried out to clone putrescine N-methyltransferase (PMT) gene which is a key enzyme in diverting polyamine metabolism towards the biosynthesis of nicotine and related alkaloids from Burley tobacco. To induce expression of PMT gene in tobacco plant, the floral meristem was removed and then mRNA was purified from root. cDNA encoding PMT gene was isolated by RT PCR and cloned. Three different groups of clones were screened by PCR and restriction enzyme digestion analysis and were characterized. The data of these screening revealed that three types of PMT are present in Burley tobacco. Comparison of the nucleotide sequence of this three genes encoding putative PMT with those of other tobaccos revealed that two types of PMT are newly discovered from Nicotiana tabacum cv. Br21 tobacco and they were same as PMT2, PMT3 of N. tabacum cv. Xanthi.

• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.32-38
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• 2021

Tubulysins are a group of secondary metabolites produced by myxobacteria that inhibit the function of the eukayotic cytoskeleton. We developed a pair of PCR primers that specifically amplified tubulysin biosynthetic genes. Using these primers, eight out of the eighty-one strains of myxobacteria belonging to the Cystobacteraceae family that harbored putative tubulysin biosynthetic genes were screened through PCR analysis. The selected strains included two Archangium gephyra, two Stigmatella sp., two Vitiosangium cumulatum, and two unidentified myxobacteria. LC-MS analysis of the culture extracts from the selected strains revealed that A. gephyra KYC4066 produced putative tubulysin A and B.

• Korean Journal of Veterinary Service
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• v.29 no.2
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• pp.103-110
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• 2006

Ticks cause economic losses to the deer industry by decreasing the growth and production of the farmed animals. The mediation of ticks affects humans and animals by causing contagious disease both directly and indirectly. Blood from farmed deer from the areas near Jangsu branch was collected for screening of infectious protozoa and rickettsial disease. Seventy deer blood samples were collected from 30 different deer farms located in Jinan, Jangsu and Muju. This blood samples were used for blood slide smear examination and hematological analysis. DNA from these samples was extracted and was used for PCR analysis for detection of gene fragments of Theileria spp, Babesia spp, Anaplasma spp and Ehrlichia spp. In the blood slide smear examination and PCR analysis all samples did not show presence of protozoal and rickettsial diseases. Eight blood samples showed anemia, 1 sample showed iron deficiency and 7 samples showed regenerative anemia. Results for PCR analysis showed 2 samples were positive for T orientalis. All DNA samples were negative for Babesia spp, Anaplasma spp, and Ehrlichia spp.

• Journal of Microbiology
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• v.45 no.1
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• pp.29-33
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• 2007

Salmonella typhimurium, causing mouse typhoid, infects hosts such as macrophage cells, and proliferates in intracellular vacuoles causing infected cells to trigger numerous genes to respond against the infection. In this study, we tried to identify such genes in RAW264.7 cells by using the PCR screening method with degenerate primers. Fourteen genes were found to be differentially expressed after a 4 h infection in which the expression of 8 genes increased while expression of the others decreased. Most of the genes were involved in proinflammatory responses such as cytokines production and cell death. The mutation in msbB gene encoding the myristoyl transferase in lipid A of lipopolysaccharide (LPS) resulted in much lower toxicity to the inoculated animals. We compared the expression of the identified genes in wild-type and msbB-mutated S. typhimurium infections and found that Lyzs encoding lysozyme type M was differentially expressed. This gene is quite likely to be related to bacterial survival in the host cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.490-496
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• 2002

In order to detect the DNA heteropolymorphism of chum salmon, selected essential genes were examined in different regional chum salmons, i.e., Korean, Japanese and American by denaturing gradient gel electrophoresis (DGGE) and real time PCR methods. From the promoter regions and introns of growth hormone, mtDNA NDI region, D-loop region, IGF-I, histone H3 and MCH2 several representative primer pairs were obtained and employed for the DGGE with the PCR products from the genomic DNAs of the different regional chum salmons. mtDNA NDI, D-loop region and IGE-I genes showed marked heteropolymorphism between Korean and American chum salmons. Intron C of growth hormone also showed a heteropolymorphism between Korean and Japanese chum salmons. Whereas heteropolnnorphism of histone liH and MCH2 genes was detected among in Korean, Japanese and Asnerican chum salmons in the examined region. The real time PCR disclosed the characteristic incremental production of target DNAs dependent on the heteropolymorphic conditions of genomic DNAa of chum salmons, thus the different regional chum salmons could be grouped by the variable incremental curies. Although the DGGE and real time PCR did not produce the identical results in this study, we suggest that the DGGE and real time PCR could be used for the primary screening of the DNA heteropolymorphism of different animal genome.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.77-81
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• 2007

Diagnostic performance of polymerase chain reaction (PCR) for detecting Dirofilaria immitis in dogs was evaluated when no gold standard test was employed. An enzyme-linked immunosorbent assay test kit (SnapTM, IDEXX, USA) with unknown parameters was also employed. The sensitivity and specificity of the PCR from two-population model were estimated by using both maximum likelihood using expectation-maximization (EM) algorithm and Bayesian method, assuming conditional independence between the two tests. A total of 266 samples, 133 samples in each trial, were randomly retrieved from the heartworm database records during the year 2002-2004 in a university animal hospital. These data originated from the test results of military dogs which were brought for routine medical check-up or testing for heartworm infection. When combined 2 trials, sensitivity and specificity of the PCR was 96.4-96.7% and 97.6-98.8% in EM and 94.4-94.8% and 97.1-98% in Bayesian. There were no statistical differences between estimates. This finding indicates that the PCR assay could be useful screening tool for detecting heartworm antigen in dogs. This study was provided further evidences that Bayesian approach is an alternative approach to draw better inference about the performance of a new diagnostic test in case when either gold test is not available.

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.255.2-255
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• 2001

No Abstract, See Full Text