• Food Science of Animal Resources
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• v.26 no.1
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• pp.121-126
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• 2006

Major milk proteins have considerable variane which comes from substitution and deletions in their amino arid sequences. Variants in genes that code for milk proteins, such as ${\beta}$-lactoglobulin (${\beta}-LG$) have been established as genetic markers for milk production and milk protein composition in dairy cattle. The effect of ${\beta}-LG$ variant on milk production traits, such as milk yield. fat yield, protein yield, fat percentage and protein percentage, was estimated for 482 Holstein cows in the first lactation. The ${\beta}-LG$ variants were determined by PCR-RFLP technique at the DNA level. Single trait linear model was used for the statistical analysis of the data. Results of this study indicated that ${\beta}-LG$ variants affected significantly protein yield (p<0.05) and fat percentage (p<0.05). Animals with the AA variant produced 31kg of milk protein more than animals with the BB variant. On the contrary, cows with the BB variant had fat percentage higher by 0.35 and 0.32% compared with cows with the AA and AB variants, respectively. No associations between the ${\beta}-LG$ variants and milk yield, protein percentage and fat yield were found Therefore, milk production traits could be improved through ${\beta}-LG$ typing by increasing the frequency of A variant for protein yield or the frequency of B variant for fat content in Holstein dairy cattle population.

• Animal cells and systems
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• v.1 no.1
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• pp.129-134
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• 1997

HLA-A2 is one of the most diversified HLA-class I antigen with 17 subtypes so far identified at the molecular level. HLA-A*02 subtyping has significant implications on the tissue typing for organ and bone marrow transplantations. Recently, DNA-based typing methods have been successfully applied to the elucidation of HLA gene polymorphisms. In the present study, HLA-A*O2 genotyping was established by using nested polymerase chain reaction-sequence specific primers (PCR-SSP) and distribution of A*O2 alleles were determined in Korean individuals. Genomic DNA prepared from four B-lymphoblastoid cell lines and lymphocytes from serologically defined 48 HLA-A2 Korean individuals by phenol/chloroform extractions was typed. The results of the four B-lymphoblastoid cells were consistent with the previous data typed by PCR analysis. Five A*O2 alleles-A*0201, A*0203, A*0206, A*0207 and A*0210-were commonly observed in a total of 17 A*02 alleles. Of these, A*0207 (f=49.0%) was the most frequent allele in Korean population. A*0206 (f=28.3%) and A*0201 (f=17.0%) were also found frequently while A*0203 and A*0210 types were observed in less than 5%. In conclusion, the high level of discrimination for HLA-A*O2 alleles will prove useful and informative in the study of transplant survival, and may identify the importance of allelic differences not readily detectable by serology on host and donor compatibility.

• Biomedical Science Letters
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• v.24 no.3
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• pp.221-229
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• 2018

Trichophyton rubrum is one of the well-known pathogenic fungi and causes dermatophytosis and cutaneous mycosis in human world widely. However, there are not an available sequence type (ST) classification methods and previous studies for T. rubrum until now. Therefore, currently, molecular biological tools using their DNA sequences are used for genotype identification and classification. In the present study, in order to characterize the genetic diversity and the phylogenetic relation of T. rubrum clinical isolates, five different housekeeping genes, such as actin (ACT), calmodulin (CAL), RNA polymerase II (RPB2), superoxide dismutase 2 (SOD2), and ${\beta}$-tubulin (BT2) were analyzed using by multilocus sequence typing (MLST). Also, DNA sequence analysis was performed to examine the differences between the sequences of Trichophyton strains and the identified genetic variations sequence. As a result, most of the sequences were shown to have highly matched rates in their housekeeping genes. However, genetic variations were found on three different positions of ${\beta}$-tubulin gene and were shown to have changed from $C{\rightarrow}G$ (1766), $G{\rightarrow}T$ (1876), and $C{\rightarrow}A$ (1886). To confirm the association with T. rubrum inheritance, a phylogenetic tree analysis was performed. It was classified as four clusters, but there was little significant correlation. Even so, MLST analysis is believed to be helpful for determining the genetic variations of T. rubrum in cases where there is more large-scale data accumulation. In conclusion, the present study demonstrated the first MLST analysis of T. rubrum in Korea and explored the possibility that MLST could be a useful tool for studying the epidemiology and evolution of T. rubrum through further studies.

• Biomedical Science Letters
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• v.19 no.3
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• pp.213-223
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• 2013

The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

• Biomedical Science Letters
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• v.23 no.3
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• pp.223-229
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• 2017

Fungal infections by human pathogenic fungi are increasing globally in elderly, children and immune suppressed or deficient patients. Aspergillus fumigatus is one of the well-known pathogenic fungi and causes aspergilloses in human world widely. However, current identification and classification methods based on its phenotypic characteristics still have limitations. Therefore, currently, molecular biological tools using their DNA sequences are used for genotype identification and classification. In the present study, in order to analyze genetic variations of A. fumigatus clinical isolates, a total of six housekeeping genes were amplified by PCR using specific primer pairs and multi-locus sequence typing (MLST) assay. Results from phylogenetic tree analysis showed that most A. fumigatus strains (88.9%) from respiratory specimens were classified into cluster A and B, and approximately half of A. fumigatus strains (46%) from non-respiratory specimens were classified into cluster C and D. Although the sample size was limited, genetic characteristics of A. fumigatus clinical isolates according to their origins were very similar and well-correlated with other clinical data.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.4
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• pp.447-452
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• 2017

In this study, we tried to identify a sea anemone collected from the coast of Gijang, Busan. The anemone was morphologically similar to species belonging to the genus Anthopleura, but its morphological characteristics did not allow for confirmed identification to species level. Multiple genes from mitochondrial cytochrome oxidase III, 12S and 16S rRNA, and nuclear 18S and 28S rRNA, were amplified for multilocus sequence typing (MLST) analysis using genomic DNA extracted from the sampled anemone and a different primer set. Based on the MLST analysis, the anemone obtained in this study was identified as Anthopleura artemisia. Also, the sequence of internal transcribed spacer-2 was most closely related to A. artemisia, indicating that this single region might be useful for anemone identification. This study shows significance of molecular identification for sea anemones, and will be helpful in studies of sea anemone identification using genotyping-by-sequencing.

• Parasites, Hosts and Diseases
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• v.52 no.3
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• pp.299-304
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• 2014

This study aimed to identify the assemblages (or subassemblages) of Giardia duodenalis by using normal or nested PCR based on 4 genetic loci: glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), ${\beta}$-giardin (bg), and small subunit ribosomal DNA (18S rRNA) genes. For this work, a total of 216 dogs' fecal samples were collected in Guangdong, China. The phylogenetic trees were constructed with MEGA5.2 by using the neighbor-joining method. Results showed that 9.7% (21/216) samples were found to be positive; moreover, 10 samples were single infection (7 isolates assemblage A, 2 isolates assemblage C, and 1 isolate assemblage D) and 11 samples were mixed infections where assemblage A was predominant, which was potentially zoonotic. These findings showed that most of the dogs in Guangdong were infected or mixed-infected with assemblage A, and multi-locus sequence typing could be the best selection for the genotype analysis of dog-derived Giardia isolates.

• Korean Journal of Organic Agriculture
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• v.12 no.4
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• pp.451-461
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• 2004

In an animals, identification system has been widely used by ear tag with dummy code and blood typing for parernity. Also, genotyping methods were using for useful mean of individual identification for live animals. In the case of genotyping estimation of gene in population of korean native chicken. In this study, we tested for development of genetic markers used it possible to determination of individual identification system. The candidate genetic markers were used already bow 10 of microstalite DNA sequence information in chromosome No. 1 and 14. Result of analysis for genotyping, the number of alleles of those microstatelites DNA was shown minimal 3 to 12 and the heterozygote expression frequency range was shown from 0.617 to 0.862. In our result, effective number of allele for each microsatellites DNA was shown 3~7, and the accuracy of individual identification was shown nearly 100%, when used with 6 genetic marker. This study was about genotyping method for identification used specific genetic marker form microsatellite DNA in the brand marketing of korean native chicken. Our results suggest that genotyping method used specific genetic marker from microsatellite DNA might be very useful for determination of individual identification.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.4
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• pp.265-272
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• 2022

The emergence and dissemination of carbapenemase-producing Enterobacteriaceae (CPE), particularly the Klebsiella pneumoniae carbapenemase-2 (KPC-2) producing Klebsiella pneumoniae, has been rapidly increasing worldwide and is becoming a serious public health threat. Since the epidemiology and characteristics of these KPC-2-producing K. pneumoniae vary according to the region and period under consideration, this study investigated the prevalence of carbapenemases and the epidemiological relationship of 78 carbapenem-resistant K. pneumoniae (CRKP) isolated from a tertiary hospital in Daejeon, from March 2017 to December 2020. The antimicrobial susceptibility tests were identified using the disk-diffusion method. PCR and DNA sequencing were used to determine the carbapenemase genes. In addition, molecular epidemiology was performed by multilocus sequence typing (MLST). Among the 78 CRKP isolates, 35 isolates (44.9%) were carbapenemase-producing K. pneumoniae (CPKP) and the major carbapenemase type was KPC-2 (30 isolates, 85.7%). The New Delhi metallo-enzyme-1 (NDM-1) and NDM-5 were identified in 4 isolates (11.4%) and 1 isolate (2.9%), respectively. Multilocus sequence typing (MLST) analysis showed 10 sequence types (STs) and the most prevalent ST was ST307 (51.4%, 18/35). All the ST307 isolates were KPC-2-producing K. pneumoniae and were multidrug-resistant (MDR). In addition, ST307 has gradually emerged during a four-year period. These findings indicate that continuous monitoring and proper infection control are needed to prevent the spread of KPC-2-producing K. pneumoniae ST307.

• Parasites, Hosts and Diseases
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• v.53 no.6
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• pp.749-753
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• 2015

Toxoplasma gondii atypical type II genotype was diagnosed in a pet peach-faced lovebird (Agapornis roseicollis) based on histopathology, immunohistochemistry, and multilocus DNA typing. The bird presented with severe neurological signs, and hematology was suggestive of chronic granulomatous disease. Gross post-mortem examination revealed cerebral hemorrhage, splenomegaly, hepatitis, and thickening of the right ventricular free wall. Histologic sections of the most significant lesions in the brain revealed intralesional protozoan organisms associated with malacia, spongiform changes, and a mild histiocytic response, indicative of diffuse, non-suppurative encephalitis. Immunohistochemistry confirmed the causative organisms to be T. gondii. DNA isolated from the brain was used to confirm the presence of T. gondii DNA. Multilocus genotyping based on SAG1, altSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico markers demonstrated the presence of ToxoDB PCR-RFLP genotype #3 and B1 gene as atypical T. gondii type II. The atypical type II strain has been previously documented in Australian wildlife, indicating an environmental transmission route.