• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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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.

• 대한의생명과학회지
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• 제26권3호
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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.

• 한국미생물·생명공학회지
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• 제18권6호
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• pp.640-646
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• 1990

E.coli의 cytidine deaminase(cytidine/2'-deoxy-cytidine aminohydrolas` EC 3.5.4.5)를 코딩하는 cdd 유전자를 E.coli cdd- pyr- 결손 변이주를 cloning host로 하여 southern blotting과 colony hybridization을 통하여 클로닝하였다. cdd 유전자가 단편인, cdd 유전자의 transcription initiation 부위의 23개 nucleotide를 합성한 후 probe로 사용하여 Southern hybridization에 의해 회수된 cdd 유전자를 함유한 단편을 얻었으며, 이를 pBR322에 삽입한 후 형질전환하여 colony hybridization한 결과 cdd+ cell을 얻었다. 삽입된 DNA 단편의 size는 27kb이었으며 이를 결실 및 subcloning을 연속 수행한 결과 2.1kb의 SalI/ DraI fragment(pTK605)에 cdd 유전자가 location 되어 있음을 알게 되었다. Mini cell 실험결과 합성된 cytidine deaminase의 활성이 pBR322에서 증폭시킴으로서 37배 정도 배가되었으며, pBR322에 비해 pUC vector계에서 다시 활성이 7배 정도 증가됨을 알 수 있었다.

• BMB Reports
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• 제39권4호
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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.

• 미생물학회지
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• 제29권5호
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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.

• 대한의생명과학회지
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• 제19권4호
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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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• 제17권6호
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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.

• 대한수의학회지
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• 제39권1호
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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.

• 한국양봉학회지
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• 제33권4호
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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.

• 한국작물학회지
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• 제48권6호
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• pp.419-423
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• 2003

우리나라 자포니카 품종의 흰잎마름병 저항성 유전자와 연관된 마커를 탐색하기 위하여, 밀양121호, 밀양123호 및 HB10624-AC5 등을 교배친으로 한 두 조합의 약배양 계통을 재료로 흰잎마름병 저항성 유전자(Xa-1 and Xa-3)와 DNA 마커간의 연관분석을 통하여 유전자 지도를 작성하고자 수행하였던 결과를 요약하면 다음과 같다. 1. $\textrm{K}_1$ 균주에 대한 흰잎마름병 저항성 검정결과, 밀양121호/HRl1650-1-4-2에서는 저항성과 감수성이 1:1로 분리하였으며, 밀양123호/HR10624-AC5 조합의 $\textrm{K}_1$ 및 $\textrm{K}_3$ 균주에 대한 검정결과는 각각 3:1과 1:1로 분리하여 이론치에 합당하였다. 2. 교배친에 대하여 DraI. HindIII, EcoRI, EcoRV, PstI등 5가지 제한효소에 대한 다형현상을 검정한 결과, RZ590, RG303, RZ536 등 3개의 마커가 다형현상을 나타내었다. 3. 흰잎마름병 포장저항성 검정결과와 RFLP 마커와의 연관분석 결과 Xa-1 유전자는 RZ590과 4번 염색체 상에서 3.1$\times$1.5 cM으로 연관되어 있었으며, Xa-3 유전자는 Rz536 및 RG303과 11번 염색체 상에서 각각 7.6$\times$2.3 및 16.0$\times$3.2 cM으로 연관되어 있었다. 4. 11번 염색체 상에서 Xa-3와 Rz536 및 RG303은 "Xa-3-RZ536-RG303" 순으로 위치하였다.순으로 위치하였다.