• Title/Summary/Keyword: Molecular genetic analysis

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A Study on the Speciadon of a Fresh Water Fish Zacco temminckL VII. Vadation of Mitochondrial DNA between 2 Types of Zacco temmincki (갈겨니(Zacco temminki)의 진화에 관한 연구 VII.갈겨니 2 Type의 Mitochondrial DNA변이)

  • Lee, Hei-Yung;Yang, Suh-Yung;Paik, Sang-Gi;Park, Chang-Shin;Yu, Sung-Lim;Lee, Sung-Keun
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.236-242
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    • 1988
  • Mitochondrial DNAs of two Mdh allelotypes of the dark chub, Z. temmincki inhabiting in Korean fresh water, were analysed. Samples of each type were collected from four populations, and the fragment patterns for mtdNA of each type were explained from 7 of the eleven restriction enzymes with hexanucleotide recognition site. Genome size was approximately 16.7 kilobases. The highly typical mtdNA fragments of each type were discovered in digestion profiles produced by Eco RI and Pst I enzyrnes. The comparisons of restriction fragment patterns and relative digestion maps permitted the estimation of fragment homology (F) and nucleotide sequence divergence(p). Between the two identical types, sequence divergence(p) was 0.128(MS), and 0.045(MM), ; between the two different types, 0.195 (range 0.177-0.226). These result may provide a distinct difference more than the value derived from allozyrne analysis, and a powerful new molecular approach for assessing genetic-evolutionary relationship among fishes.

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A Novel RUNX2 Mutation in a Korean Family with Cleidocranial Dysplasia (한국인 쇄골 두개 이형성증 가족에서의 RUNX2 유전자 돌연변이)

  • Lee, Ji Won;Song, Jisoo;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Young-Jae;Lee, Sang-Hoon;Kim, Jongbin;Kim, Jung-Wook
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.4
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    • pp.409-415
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    • 2019
  • Cleidocranial dysplasia (CCD) is an autosomal-dominant disease characterized by the delayed closure of cranial sutures, defects in clavicle formation, supernumerary teeth, and delayed tooth eruption. Defects in the Runt-related transcription factor 2 (RUNX2), a master regulator of bone formation, have been identified in CCD patients. The aim of this study was to identify the molecular genetic causes in a CCD family with delayed tooth eruption. The 23-year-old female proband and her mother underwent clinical and radiographic examinations, and all coding exons of the RUNX2 were sequenced. Mutational analysis revealed a single nucleotide deletion mutation (NM_001024630.4 : c.357delC) in exon 3 in the proband and her mother. The single C deletion would result in a frameshift in translation and introduce a premature stop codon [p.(Asn120Thrfs*24)]. This would result in the impaired function of RUNX2 protein, which may be the cause of delayed eruption of permanent teeth in the family.

Molecular Genetic Classification of Hypsizigus marmoreus and Development of Strain-specific DNA Markers (느티만가닥버섯의 분자유전학적 분류 및 품종특이적 DNA 마커 탐색)

  • Lim, Yun-Jeong;Lee, Chang-Yun;Park, Jeong-Eun;Kim, Sang-Woo;Lee, Hyun-Sook;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.34-39
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    • 2010
  • We have attempted to verify 30 strains of Hypsizigus marmoreus from various mushroom stocks in Korea using random amplified polymorphic DNA (RAPD) methodology. Chromosomal DNAs of them were extracted and subjected to PCR analyses with 3 random primers. Each PCR produced approximately 30 distinct PCR bands with the size from 200 bp to 3000 bp. A dendrogram was acquired using the unweighted pair-group method with arithmetic average (UPGMA) clustering methodology on the basis of the DNA band pattern. The analysis revealed that 30 strains of H. marmoreus were clustered into two distinct clusters. Cluster 1 contained 3 subgroups while the cluster 2 consisted of rather diverse strains. Interestingly, Hm3-10, a wild strain collected from Deog-Yu mountain, was not included in either clusters, indicative of uniqueness of this strain. We nextly attempted to develop strain-specific DNA markers to verify a specific strain. A unique band in the RAPD gel lane of Hm0-4 was extracted and its sequence was determined. PCR with a primer set from the determined sequence revealed that the primer set gave a 250 bp DNA band only for Hm0-4, indicating that this approach works well for the strain-specific identification of H. marmoreus.

Mapping QTLs for Agronomic Traits Using an Introgressin Line Population from a Cross between Ilpumbyeo and Moroberekan in Rice (일품벼/모로베레칸 이입계통을 이용한 농업형질 관련 QTL 분석)

  • Ju, Hong-Guang;Kim, Dong-Min;Kang, Ju-Won;Kim, Myung-Ki;Kim, Yeon-Gyu;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.40 no.4
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    • pp.414-421
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    • 2008
  • We conducted a QTL analysis of agronomic traits using 117 $BC_3F_5$ and $BC_3F_6$ lines developed from a cross between Ilpumbyeo and Moroberekan. Genotypes of 117 $BC_3F_5$ lines were determined using 134 simple sequence repeat (SSR) markers. A total of 832 Moroberekan chromosome segments with 410 homozygous and 422 heterozygous, respectively, were detected, and the genetic distance of introgression segments ranged from 0.5 cm to 112.1 cm. A linkage map constructed using 134 SSR markers was employed to characterize quantitative trait loci (QTL). The 117 $BC_3F_5$ and $BC_3F_6$ lines were evaluated for seven agronomic traits at two locations in 2006 and 2007 and at one location in 2007. A total of 26 QTLs were identified for seven traits including days to heading, and the phenotypic variance explained by each QTL ranged from 9.2% to 24.2%. Moroberekan alleles contributed positive effects in the Ilpumbyeo background at eleven QTL loci including panicle length and spikelets per panicle. Five QTLs, two for days to heading and one each for culm length, panicle length and spikelets per panicle were consistently detected in every occasions indicating that these QTLs are stable. Among them, two QTLs, spp6 for spikelets per panicle and pl6 for paniclel length were localized in the similar region. Increase in spikelets per panicle at this locus might be due to the increase in panicle length, because both traits were associated with increase in spikelets per panicle and panicle length due to the presence of the Moroberekan allele. These Moroberekan QTLs might be useful in breeding programs to develop high-yielding cultivars.

Inheritance of P34 Allergen Protein in Mature Soybean Seed

  • Sung, Mi Kyung;Seo, Jun Soo;Kim, Kyung Roc;Han, Eun Hui;Nam, Jin Woo;Kang, Dal Soon;Jung, Woo Suk;Kim, Min Chul;Shim, Sang In;Kim, Kyung Moon;Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.43 no.2
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    • pp.115-119
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    • 2011
  • Soybean proteins are widely used for human and animal feeds worldwide. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are present in the raw mature soybean. P34 protein, referred as Gly m Bd 30K, has been identified as a predominant immunodominant allergen. The objective of this research is to identify the genetic mode of P34 protein for the improvement of soybean cultivar with a very low level of P34 protein. Two $F_2$ populations were developed from the cross of "Pungsannamulkong" ${\times}$ PI567476 and "Gaechuck2ho" ${\times}$ PI567476 (very low level of P34 protein). Relative amount of P34 protein was observed by Western blot analysis. The observed data for the progeny of "Pungsannamulkong" and PI567476 were 133 seeds with normal content of P34 protein and 35 seeds with very low level of P34 protein (${\chi}^2=1.157$, P=0.20-0.30). For the progeny of "Gaechuck#1" and PI567476, the observed data were 177 seeds with normal content of P34 protein and 73 seeds with very low level of P34 protein (${\chi}^2=2.353$, P=0.10-0.20). From pooled data, observed data were 310 seeds with normal content of P34 protein and 108 seeds with very low level of P34 protein (${\chi}^2=0.156$, P=0.50-0.70). The segregation ratio (3:1) and the Chi-square value obtained from the two populations suggested that P34 protein in mature soybean seed is controlled by a single major gene. Single gene inheritance of P34 protein was confirmed in 32 $F_2$ derived lines in $F_3$ seeds, which were germinated from the low level of P34 protein obtained from the cross of "Pungsannamulkong" and PI567476. These results may provide valuable information to breed for new soybean line with low level of P34 protein and identification of molecular markers linked to P34 locus.

Characteristics of Viral Endothelial Cell Necrosis of Eel (VECNE) from Culturing Eel (Anguilla japonica, Anguilla bicolar) in Korea (국내 양식산 뱀장어(Anguilla japonica, Anguilla bicolar)에서 발생하는 바이러스성내피세포괴사증(VECNE)의 감염 특성)

  • Kim, Su Mi;Ko, Sang Mu;Jin, Ji Hye;Seo, Jung Soo;Lee, Nam Sil;Kim, Young Suk;Gu, Jeong Hui;Bae, Yu Ri
    • Korean Journal of Ichthyology
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    • v.30 no.4
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    • pp.185-193
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    • 2018
  • From 2017 to 2018, the disease has been monitored at four culturing eel farms in Incheon and Gyeonggi region in Korea. As a result, diseases with gill congestion frequently occurred. This disease occurred regardless of size of eel, but the frequency and cumulative mortality were high in eels within 3 months after stocking. The infected fish showed pathological histopathological features such as intense congestion and dilation in the central venous sinus (CVS) of gill filaments and hemorrhages in liver and kidney. Hexagonal viral particles measuring about 70 nm in diameter was observed in nuclei and cytoplasm of gill vascular endothelial cells. Molecular biologic diagnosis by both PCR and genetic analysis has been revealed that the causative agent of this disease is Japanese eel endothelial cells-infecting virus (JEECV), the cause of viral endothelial cell necrosis of eel (VECNE), which is mainly reported in Japan. This study is the first report on the characteristics of JEECV and VECNE infection in domestic eel farms.

Characterization of Phenotypic Traits and Application of Fruit Flesh Color Marker in Melon (Cucumis melo L.) Accessions (멜론 유전자원의 생육 평가와 과육색 유전형 분석)

  • Bae, Ik Hyun;Kang, Han Sol;Jeong, Woo Jin;Ryu, Jae Hwang;Lee, Oh Hum;Chung, Hee
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.478-490
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    • 2021
  • We aimed to generate basic breeding data for melon (Cucumis melo L.). A total of 219 melon accessions conserved at the National Agrobiodiversity Center (NAC) in Rural Development Administration (RDA) were used in this study, of which 72 (33%) were collected from India. The majority of accessions showed orange (42%) and white (36%) flesh color. In addition to phenotypic evaluations, the accessions were genotyped using a molecular marker for the carotenoid biosynthesis gene CmOr. DNA fragments of the expected size were amplified in 205 out of 219 accessions. Digestion of the PCR products with HinfI restriction endonuclease showed 100% concordance between phenotype and genotype in green-fleshed accessions, but 98%, 97%, and 80% concordance in orange-, white-, and creamy-fleshed accessions, respectively. Sequence analysis revealed single nucleotide changes in the three positions of SNP1, SNP2 and SNP1int in the CmOr gene among accessions. These newly found alleles suggest that there are multiple mechanisms in determining fruit flesh color in melon. Also, the phenotype data of diverse accessions obtained in this study will be a valuable source for melon breeding.

Isolation and identification of tick-borne pathogens in hard ticks collected in Daejeon (대전 주택가 산책로 진드기의 인수공통전염병 병원체 감염실태 조사)

  • Han, So-young;Sung, Sun-hye;Seo, Jin-woo;Kim, Jong-ho;Lee, Seok-ju;Yoo, Sang-sik
    • Korean Journal of Veterinary Service
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    • v.44 no.2
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    • pp.93-102
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    • 2021
  • In this study, a total of 9,449 hard ticks were collected once a month from April to October 2020 from a neighborhood park in Daejeon by flagging & dragging method and CO2 manned trap method. The collected ticks were classified according to the Yamagutsi search table using a stereoscopic microscope and molecular biological analysis of four pathogens (SFTSV, Anaplasma spp., Ehrlichia spp., Borrellia spp.). As a result of the study, Haemaphysalis longicornis were collected the most in all areas of the five boroughs at a rate of 82 to 96 percent, while adults were collected the most in May to July, nymphs were collected the most in April to June, and larvae from August to October at a rate of 78 percent to 98 percent. In pathogens, three cases of SFTSV were detected, showing a minimum infection rate (MIR) of 0.46%, while Anaplasma spp. and Ehrlichia spp. were detected one each, with 0.15% and Borrelia spp. with a minimum infection rate of 0.46%. The detected SFTSV showed 99.9% homogeneity with the KF781490 detected in Cheongwon-gun, Chungbuk Province, Anaplasma spp. showed 99.0% homogeneity with JN990105 detected in China, and Erhlichia spp. showed 98.9% genetic similarity with U96436 separated from the U.S. In this study, the distribution status and pathogen infection rate of the hard ticks in the Daejeon area are analyzed and provided as basic data for the prevention of the hard tick-borne infectious disease.

Detection of the Carbapenem Resistance Gene in Gram-negative Rod Bacteria Isolated from Clinical Specimens (임상검체에서 분리된 그람음성막대균으로부터 카바페넴 내성 유전자 검출)

  • Yang, Byoung Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.3
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    • pp.179-191
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    • 2022
  • Carbapenem-resistant Enterobacteriaceae (CRE) poses an increasing public health threat and has limited treatment options with high associated mortality. Genotypes of carbapenemase that threaten public health (blaKPC, blaNDM, blaIMP, and blaVIM) and blaOXA-48-like genes were detected by phenotypic and molecular diagnosis, and related gene distribution patterns were investigated. Phenotypic testing using the modified Hodge test confirmed positivity in all 41 strains examined, and carbapenemase inhibitory testing using meropenem+phenyl boronic acid or meropenem+EDTA confirmed positivity in 18 and 8 strains, respectively. Polymerase chain reaction revealed the presence of amplification products in 28 strains of blaKPC, 25 strains of blaNDM, 5 strains of blaIMP, 1 strain of blaVIM, and 13 blaOXA-48-like strains. In addition, 7 strains of blaKPC+blaNDM, 1 strain of blaKPC+blaIMP, 1 strain of blaNDM+blaOXA-48-like, 1 strain of blaNDM+blaVIM, 4 strains of blaKPC+blaNDM+blaIMP, and 4 strains of blaKPC+blaNDM+blaOXA-48-like were identified. Melting curve analysis using real-time PCR was wholly consistent with PCR results. The study shows genetic identification of highly specific CRE by real-time PCR could be used to provide early diagnoses and infection control, improve surveillance, and prevent the transmission of CRE.

The List of Korean Organisms Registered in the NCBI Nucleotide Database for Environmental DNA Research (환경유전자 연구를 위한 NCBI Nucleotide 데이터베이스에 등록된 국내 생물 목록 현황)

  • Ihn-Sil Kwak;Chang Woo Ji;Won-Seok Kim;Dongsoo Kong
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.352-359
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    • 2022
  • Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.