• Asian-Australasian Journal of Animal Sciences
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• v.27 no.9
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• pp.1228-1235
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• 2014

The swine leukocyte antigen (SLA)-DRA locus is noteworthy among other SLA class II loci for its limited variation and has not been investigated in depth. This study was investigated to detect polymorphisms of four exons of SLA-DRA gene and its association with piglet diarrhea in Landrace, Large White and Duroc pigs. No polymorphisms were detected in exon 3, while 2 SNPs (c.178G>A and c.211T>C), 2 SNPs (c.3093A>C and c.3104C>T) and 5 SNPs (c.4167A>G, c.4184A>G, c.4194A>G, c.4246A>G and c.4293G>A) were detected in exon 1, exon 2 and exon 4 respectively, and 1 SNP (c.4081T>C) in intron 3. Statistical results showed that genotype had significant effect on piglet diarrhea, individuals with genotype BC had a higher diarrhea score when compared with the genotypes AA, AB, AC and CC. Futhermore, genotype AC had a higher diarrhea score than the genotype CC in exon 1 (p<0.05); diarrhea scores of genotype AA and BB were higher than those of genotypes AC and CC in exon 2 (p<0.05); individuals with genotype AA had a higher diarrhea score than individuals with genotype AB and BB in exon 4 (p<0.05). Fourteen common haplotypes were founded by haplotype constructing of all SNPs in the three exons, its association with piglet diarrhea appeared that Hap2, 5, 8, 10, and 14 may be the susceptible haplotypes and Hap9 may be the resistant haplotype to piglet diarrhea. The genetic variations identified of the SLA-DRA gene may potentially be functional mutations related to piglet diarrhea.

• Biomedical Science Letters
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• v.26 no.3
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• pp.238-243
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• 2020

Tuberculosis is a global public health problem and manifests itself as a difference in the genetic susceptibility of the host, along with the properties of Mycobacterium tuberculosis (MTB). The single nucleotide polymorphisms (SNPs) and candidate genes proposed in the Genome-wide association study (GWAS) on tuberculosis in a recently published Chinese population were reported. In this study, we investigated whether the genetic polymorphism of candidate genes related to tuberculosis is reproduced when targeting Koreans. The CLCN6 (rs12404124, rs198391, rs535107), DOK7 (rs1203104, rs1203103) and HLA-DRA (rs1051336) gene polymorphisms showed statistically significant results. In addition, it was also found whether it acts as an expression quantitative trait loci (eQTL) that can influence gene expression. This study confirmed that the genetic polymorphism of the three genes (CLCN6, DOK7, HLA-DRA) affects the development of tuberculosis and will help to understand the genetic specificity of tuberculosis and the interaction between pathogens and hosts.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.640-646
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• 1990

We have cloned the cdd gene from E. coli C600 using (cdd-) as a host. From the sequenced promoter region of E=, coli cdd gene which has been determined by Valentin-Hansen P. (1985), we synthesized the 23 mer oligonucleotides corresponding to the transcription initiation region and used as a probe for cloning of the cdd gene by Southern blotting. The isolated fragments in the blotting were introduced to the colony hybridization after transforming it into the E. coli JF611 (cdd-, pyr double mutant), and we identified the hybridized band at 27 kb long. From the original insert of 27 kb fragment in theBamHI site of pBR322, the 5.3 kb fragment containing the cdd gene was isolated by subsequent deletion and subeloning. From the derived plasmid pTK509, further deletion and subcloning were performed and clarified that the cdd gene was located in the 2.1 kb of SaZI/DraI segment in the insert of pTK605. The polypeptide encoded by the cloned DNA was appeared to be a molecular mass of 33,000.

• BMB Reports
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• v.39 no.4
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• pp.418-425
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• 2006

A human bacterial artificial chromosome (BAC) library was constructed with high molecular weight DNA extracted from the blood of a male Korean. This Korean BAC library contains 100,224 clones of insert size ranging from 70 to 150 kb, with an average size of 86 kb, corresponding to a 2.9-fold redundancy of the genome. The average insert size was determined from 288 randomly selected BAC clones that were well distributed among all the chromosomes. We developed a pooling system and three-step PCR screen for the Korean BAC library to isolate desired BAC clones, and we confirmed its utility using primer pairs designed for one of the clones. The Korean BAC library and screening pools will allow PCR-based screening of the Korean genome for any gene of interest. We also determined the allele types of HLA-DRA and HLA-DRB3 of clone KB55453, located in the HLA class II region on chromosome 6p21.3. The HLA-DRA and DRB3 genes in this clone were identified as the DRA*010202 and DRB3*01010201 types, respectively. The haplotype found in this library will provide useful information in future human disease studies.

• Korean Journal of Microbiology
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• v.29 no.5
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• pp.290-295
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• 1991

N4 phage, which infects E. coli K-12 strains, could not infect E. coli K-12 strains containing rtn(resistant to N4) gene on plasmids, which was isolated from Proteus vulgaris ATCC 13315. The region of rtn gene for Rtn phenotype was reduced to the 1.7 kb HincII-AccI fragment, and rtn gene seemed to have its own promoter. This putative promoter was present in 107 bp HindII-DraI fragment, and known to be functional in E. cole K-12, which is supported by the fact that phenotype of a subclone, pRMG103A1B which does not contain the 107 bp fragment, was dependent on the existance of a functional promoter in the upstream of rtn gene, and that the 107 bp fragment had promoter activity when located in the upstream of structural gene of galactodinase of E. coli. The promoter-bearing fragment contains two overlapping putative promoter sequences, both of which show a fit in eight of twelve nucleotides with consensus sequences of E. coli promoters at the -35 and -10 regions.

• Biomedical Science Letters
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• v.19 no.4
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• pp.310-314
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• 2013

CYP2E1 in the liver has been studied intensively because it is involved in the metabolic activation of xenobiotics. It is inducible by alcohol, so it has been suspected as the cause of cancer in the stomach and lung. The possible role of CYP2E1 has been suggested strongly as causing tissue damage in mice with renal failure. It was also suspected that 5'-flanking region of CYP2E1 gene might be involved with renal failure. So, we investigated polymorphism of restriction enzyme sites within CYP2E1 gene using the PCR-RFLP analysis. PstI and RsaI sites were located at 5'-flanking region and DraI site was located at intron 6. All three types (W/W, W/S, S/S) were observed for these enzymes although each incidence was somewhat different depending the enzyme sites. W/W was prominent for PstI whereas W/S was markedly high for RsaI. Overall, polymorphic incidence in patients was somewhat higher than normal population. This research should facilitate further investigation of CYP2E1 at genetic level as the direct cause of tissue damage in various organs.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.743-749
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• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.90-95
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• 1999

A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

• Journal of Apiculture
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• v.33 no.4
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• pp.227-237
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• 2018

The majority parasitic bee mites of Thailand in genus Tropilaelaps are infesting colonies of native bees (Apis dorsata) and introduced bees (A. mellifera). The investigation aims to study morphological and genetic variation of Tropilaelaps mites infected different hosts. Adult mites were collected from honey bee brood throughout Thailand. Traditional and geometrical morphometrics were measured on photograph by using TPS program. Additional, COI gene variations were examined by PCR-RFLP and nucleotides sequencing. Tree of mites relationships were constructed by NJ and MP assumptions. Morphometric results indicated T. mercedesae were major species infesting on A. dorsata and A. mellifera. Mophological variation represented at anal and epigynial plate, which the shape of the anal plate apex margin has been key character to identify between T. mercedesae (bell to blunt shape) and T. koenigerum (pear shape). However, the discriminant analysis suggested that geometric results were potential to classify Thai Tropilaelaps populations from different hosts better than traditional morphometric. Otherwise, PCR-RFLP clearly detected the site of Dra I and Xba I digestion of Thai Tropilaelaps morphotypes. The COI sequences of T. koenigerum were founded infesting only A. dorsata in Thailand and four sequences that related to the Thai T. mercedesae morphotypes. The NJ and MP tree were clearly classified Thai Tropilaelaps species which were suggested both from morphological and molecular analysis. This information might be basically of taxonomic status, but this should have implication for controlling these mites in Thailand and other countries.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.6
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• pp.419-423
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• 2003

Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.