• Korean Journal of Microbiology
• /
• v.54 no.1
• /
• pp.31-37
• /
• 2018

Campylobacter jejuni is an important food-borne pathogen causing gastroenteritis in human. We isolated 208 strains of Campylobacter jejuni from 430 diarrhea patients and food employees with 17 food-poisoning outbreaks between 2014 and 2016 in Gyeonggi area. The strains were tested for genetic relationship and the genotype distribution using PFGE and multiplex-PCR typing. Among the 47 Penner-serotypes known for C. jejuni, it was identified as a genotype consisting of 35 genotypes by multiplex-PCR typing and represented 7 genotypes (HS2, HS4A, HS8, HS15, HS29, HS41, and HS53) in the selected strain. From the PFGE analysis of 11 food-poisoning outbreaks, 5 group of PFGE profile were obtained, and genetic similarity in these clusters ranged from 61.8 to 66.6%. This study examines the genetic diversity of C. jejuni that have been separated in the Gyeonggi area through various genetic analysis methods and identifies the correlation between strains in patients who have been infected with the disease in the future.

• Korean Journal of Breeding Science
• /
• v.40 no.4
• /
• pp.387-393
• /
• 2008

Over 7 individual rice (Oryza sativa L.) plants per a line were sowed and sampled by pooled sampling method for genomic DNA extraction. The 5,400 flanking sequence tags (FSTs) were analysed by adaptor PCR and direct sequencing. FST analysis showed that the intragenic FSTs, the intergenic FSTs, and the original insertional sequences including hot spot covered 48.1% (2,597), 25.6% (1,383), and 25% (1,350), respectively. The 2,597 intragenic FSTs were used for genotyping to determine whether they are heterozygous or homozygous, and 1,393 core lines were selected. Among them, 422 knockout genes were distributed on chromosome 3, while 56 - 157 intragenic FSTs scattered on other chromosomes. Among 1,393 FSTs, known genes such as transcription factor covered 59.4% (827), while unknown genes such as expressed protein covered 40.6% (566). RT-PCR indicated that some core lines had no expression or decreased expression level in their knockout genes. It means that core lines are very useful knockout lines for functional genomic studies.

• Korean Journal of Veterinary Service
• /
• v.42 no.1
• /
• pp.1-7
• /
• 2019

Porcine reproductive and respiratory syndrome (PRRS) is economically the most important and challenging disease in swine industries worldwide and caused by PRRS virus (PRRSV). Previous studies reported that pigs with heterozygous genotypes in the guanylate-binding proteins (GBP1 and GBP5) exhibited increased resistance against PRRSV infection. The present study was conducted to produce higher numbers of the heterozygous pigs based on the PRRS resistant polymorphisms found in GBP1 (GBP1E2 and WUR) and GBP5, and evaluate the resistance of heterozygous pigs against challenge with a type 2 PRRSV (JA142) in comparison with homozygous pigs. In the challenge study, 12, 4 week-old PRRSV-negative piglets were selected based on the genotypes of the 3 polymorphisms (GBP1E2, WUR and GBP5). Among them, 8 piglets [homozygous (n=4) and heterozygous (n=4)] were challenged with JA142 and kept in the same room, and the remaining 4 piglets were kept separately as a negative control. In results, the sperms collected from the boars of GBP1E2-GG genotype produced approximately 28~41% higher numbers of heterozygous piglets as compared with those from the boars of GBP1E2-AG genotype. In the challenge study, we found that heterozygous piglets showed the significantly lower levels of viremia than homozygous piglets at 14, 21 and 28 dpc. Consistently, these heterozygous piglets also exhibited significantly higher ADWG than homozygous piglets. Therefore, in the current study, selection of boars based on SNP markers could increase the production of PRRS resistant piglets and the PRRS resistant pigs were found to be more resistant to PRRSV infection.

• IMMUNE NETWORK
• /
• v.3 no.3
• /
• pp.219-226
• /
• 2003

Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.16 no.4
• /
• pp.724-728
• /
• 2002

Angiotensin-converting enzyme (ACE) gene polymorphism, which consists of presence (insertion, I) or absence (deletion, D) of a 250-bp fragment, is associated with ischemic heart disease, renovascular disease, systemic lupus erythematosus. Subjects with the DD genotype have higher levels of circulating ACE than subjects with the II genotype and show an increased tendency towards vascular wall thickness and contribute to the development of vascular disease. But the association between I/D polymorphism of the ACE gene and cerebrovascular disease is still controversial. The aim of this study was to determine whether the DNA polymorphism of the ACE are associated with cerebrovascular disease in Korean population. The study group comprised 377 Korean patients admitted to Kyunghee Oriental Medical Center in the year of 2000 for the treatment of brain infarction or brain hemorrhage. Magnetic resonance imaging(MRI) was performed for each patient to determine the stroke phenotype, infarction or hemorrhage. The 183 subjects without evidence of brain infarction or brain hemorrhage were selected from the some ethnical population(control group). Venous blood samples were drawn from each subject for the extraction of DNA. Genotypes of ACE were determined by polymerase chain reaction amplification of the genomic DNA. Case and control genotype frequencies were compared by chi-square testing. Both the patients and the controls were classified respectively into 4 groups: age less than forty years, age forty one to fifty, age fifty one to sixty, age greater than sixty years. There were no significant differences in the distributions of ACE genotypes among the patients with infarction, with hemorrhage and controls (Infarction: D/D 15.8%, I/D 46.7%, I/I 37.5%, Hemorrhage: D/D 15.1%, I/D 46.5%, I/I 38.4%, Control: D/D 18.6%, I/D 50.3%, I/I 31.2%). There was a significant difference in the distribution of ACE genotypes between the age greater than sixty year subgroup of patient with brain hemorrhage and the control (Hemorrhage: D/D 0%, I/D 55.6%, I/I 44.4%, Control: D/D 13.0%, I/D 63.0%, I/I 23.9%; Pearson Chi-Square value 5.956, P<0.05). Furthermore, the frequency of the ACE D/D type declined with increasing age both in the patient and control group (Patient group: age < 50 D/D 21.5%, age > 50 D/D 14.42%; Control group: age < 50 D/D 21.0%, age > 50 D/D 14.2%). In conclusion there is no clear association between ACE polymorphism and cerebrovascular disease in Korean population. Although, there was a tendency for the frequency of the ACE D/D type declined with increasing age in both patients and controls.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.8
• /
• pp.1169-1173
• /
• 2007

The protein encoded by SLC27A2 gene is an isozyme of long-chain fatty-acid-coenzyme A ligase family, and it converts free long-chain fatty acids into fatty acyl-CoA esters, and thereby plays a key role in lipid biosynthesis and fatty acid degradation. In the present study, SLC27A2 located on human chromosome 15 was selected as candidate gene and we isolated and cloned partial fragments of mRNA sequence and genomic fragments of porcine SLC27A2 gene. The coding region of the gene as determined by alignments shared 90% and 82% identity with human and mouse cDNAs, respectively. Detection in LargeWhite and Meishan breeds showed that a single nucleotide polymorphism (SNP) ($A{\rightarrow}G$) existed in exon 7, which caused corresponding amino acid changed for encoding. In LargeWhite pigs it encoded for Val while in Meishan pigs it encoded for Ile, so we developed the PCR-RFLP genotype method for detection of this polymorphism. Association study in 135 $F_2$ reference family indicated that significant correlation existed between the polymorphism and growth and carcass traits.

• Korean Journal of Plant Resources
• /
• v.25 no.6
• /
• pp.762-772
• /
• 2012

The objective of this study was to analyze the genetic diversity using SSR marker and investigate the fatty acid composition of perilla (P. frutescens var. frutescens) germplasm. Genetic diversity among 95 accessions, which consisted of 29 weedy types and 66 landrace accessions, was evaluated based on 12 SSR markers carrying 91 alleles. The mean values of observed ($H_O$) and expected heterozygosities ($H_E$) were 0.574 and 0.640, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into two distinct groups, which were the landrace, moderate and weedy type, genetic distance (GD) value was 0.609. The physicochemical traits about crude oil contents and fatty acid compositions were analyzed using GC. Among tested germplasm, the total average oil contents (%) showed a range from 28.57 to 49.67 %. Five fatty acids and their contents in the crude oils are as follows: ${\alpha}$-linolenic acid (41.12%-51.81%), linoleic acid (15.38%-16.43%), oleic acid (18.93%-27.28%), stearic acid (2.56%-4.01%), and palmitic acid (7.38%-10.77%). The average oil content of wild types was lower than landrace, and the oil content of middle genotype accessions was higher than other germplasm, but no significant variation between landrace and wild types was shown. Nevertheless, IT117174, landrace of Korea, was highest in crude oil content (47.11%) and linolenic acid composition (64.58%) among the used germplasm. These traits of the selected accessions will be helped for new functional plant breeding in perilla crop.

• Horticultural Science & Technology
• /
• v.33 no.3
• /
• pp.396-402
• /
• 2015

This study identified a new male-sterile germplasm of Chinese jujube, named male-sterile No. 2 (JMS2), and achieved controlled hybridization using that germplasm as the female parent. The anthers of JMS2 before flower bud opening became shrunken, dingy yellow and much smaller than normal ones, and they changed to brown after anthesis. No pollen was observed in anthers of JMS2 and its male-sterile trait remained stable over different years. A total of 1,642 fruits were obtained from ten intra- and interspecific cross combinations via controlled hybridization from 2008 to 2012 using JMS2 as the female parent. Of those, 27.3% produced seeds, on average (0-72.6%). The rate of fruit with seed (RFS) was significantly different between cross combinations or years. Compared to other cross combinations, the RFS in the combination of JMS2 ${\times}$ 'Xingguang' (a Chinese jujube cultivar with high resistance to jujube witches' broom disease) and JMS2 ${\times}$ 'Xing16' (a sour jujube genotype) remained high in different years and reached means of 48.7 and 58.1%, respectively. Finally, 150 plantlets were regenerated from immature embryos, and 51 of them were randomly selected and identified to be authentic hybrids using amplified fragment length polymorphism (AFLP) markers. This is the first report of hybrids obtained from a cross between Chinese jujube and sour jujube.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.1
• /
• pp.41-48
• /
• 2012

Introduction: Breast cancer is the second leading cancer in Korean women. To assess potential genetic associations between the prostate stem cell antigen (PSCA) gene in the chromosome 8q24 locus and breast cancer risk in Korean women, 13 SNPs were selected and associations with breast cancer risk were analyzed with reference to hormone receptor (HR) and menopausal status. Methods:We analyzed DNA extracted from buffy coat from 456 patients and 461 control samples, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) based upon region-specific PCR followed by allelespecific single base primer extension reactions. Risks associated with PSCA genotypes and haplotypes were estimated with chi-square test (${\chi}^2$-test), and polytomous logistic regression models using odds ratios (OR) and 95% confidence intervals (CIs), by HR and menopausal status. Results: In case-control analysis, odds ratios (OR) of rs2294009, rs2294008, rs2978981, rs2920298, rs2976395, and rs2976396 were statistically significant only among women with estrogen receptor (ER) negative cancers, and those of rs2294008, rs2978981, rs2294010, rs2920298, rs2976394, rs10216533, and rs2976396 were statistically significant only in pre-menopausal women, and not in postmenopausal women. Risk with the TTGGCAA haplotype was significantly elevated in ER (-) status (OR= 1.48, 95% CI= 1.03~2.12, p<0.05). Especially risk of allele T of rs2294008 is significantly low in pre-menopausal breast cancer patients and AA genotype of rs2976395 in ER (-) status represents the increase of OR value. Conclusion: This report indicated for the first time that associations exist between PSCA SNPs and breast cancer susceptibility in Korean women, particularly those who are pre-menopausal with an estrogen receptor negative tumor status.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.58 no.2
• /
• pp.91-106
• /
• 2013

Exserohilum turcicum is considered serious destructive disease of maize (Zea mays L.) in North Korea. This study aimed to understand genetic inheritance and combining ability of newly bred lines of maize tolerant to E. turcicum by diallel crosses. Three diallel sets for two different ecological regions and one agronomic trait; eastern (E), northern (N) and stay green (SG) involving 29 inbred lines were tested in eight locations of 2000 and 2001. E. turcicum infections were under natural conditions, respectively. Lines used were selected for high yield potential in test crosses with good agronomic traits and tolerance to biotic and abiotic stresses. Selection for race specific high resistance to biotic stresses was avoided to select quantitatively inherited genes. Host plant responses to E. turcicum were rated on a scale of 1 (highly tolerant) to 9 (highly susceptible). Highly significant variations were recorded in all trials. General combining ability (GCA) mean square was roughly twice that of specific combining ability (SCA). The genotype (G) by environment (E) interaction was highly significant. The overall results of genetic studies in three diallel sets show that genetic control for inbred tolerance to E. turcicum is polygenic and quantitatively inherited. New inbreds; E-3, N-1 and SG-4 confer better tolerance to E. turcicum than the widely used inbreds; Mo17, and B73. Proper use of genetic information from this study shall increase of corn production under high E. turcicum infection in the Far Eastern Regions of Korea and China.