• 제목/요약/키워드: chromosome isolation

검색결과 59건 처리시간 0.025초

Variation of Univariate Flow Karyotypes and Chromosomal DNA Contents in Maize (Zea mays L.)

  • Lee, Jai-Heon;Lee, Myoung-Hoon;Kim, Kyung-Je
    • 한국작물학회지
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    • 제43권2호
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    • pp.128-133
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    • 1998
  • Analyses of now karyotypes using different maize (Zea mays L.) inbred lines have been performed. The accumulation and isolation of high quality and quantity metaphase chromosomes from root tips can be achieved from many kinds of maize lines. The chromosome suspensions were prepared by a simple slicing method from synchronized maize root tips and analyzed with a now cytometry. The variations of experimental now karyotypes were detected among inbred lines in terms of the positions and/or the numbers of chromosome peaks. The 2C DNA amount among 8 inbred lines ranged from 5.09 to 5.52 pg. The variability of DNA content in maize chromosome 1 was 9.1 % ranging from 0.685 to 0.747 pg. The selection of appropriate maize lines is critical for sorting specific single chromosome types. At least five different chromosome types can be discriminated and sorted from five maize lines.

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Construction of Chromosome-Specific BAC Libraries from the Filamentous Ascomycete Ashbya gossypii

  • Choi Sang-Dun
    • Genomics & Informatics
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    • 제4권2호
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    • pp.80-86
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    • 2006
  • It is clear that the construction of large insert DNA libraries is important for map-based gene cloning, the assembly of physical maps, and simple screening for specific genomic sequences. The bacterial artificial chromosome (BAC) system is likely to be an important tool for map-based cloning of genes since BAC libraries can be constructed simply and analyzed more efficiently than yeast artificial chromosome (YAC) libraries. BACs have significantly expanded the size of fragments from eukaryotic genomes that can be cloned in Escherichia coli as plasmid molecules. To facilitate the isolation of molecular-biologically important genes in Ashbya gossypii, we constructed Ashbya chromosome-specific BAC libraries using pBeloBAC11 and pBACwich vectors with an average insert size of 100 kb, which is equivalent to 19.8X genomic coverage. pBACwich was developed to streamline map-based cloning by providing a tool to integrate large DNA fragments into specific sites in chromosomes. These chromosome-specific libraries have provided a useful tool for the further characterization of the Ashbya genome including positional cloning and genome sequencing.

갈겨니 (Zacco temmincki)의 進化에 관한 硏究 II: 갈겨니 2型의 核型分析

  • 이혜영;조정우;양서영
    • 한국동물학회지
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    • 제29권3호
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    • pp.208-214
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    • 1986
  • 電氣泳動法에 의해 $Mdh-1^{MM}, Mdh-1^{MS}$ en allotype으로 분류된 경상남도 남해군 동천천産 갈겨니 (Zacco temmincki)의 核型을 비교하였다. 이들 두型의 염색체수는 2n=48로 동일하지만, 7번째 염색체는 $Mdh-1^{MM}$型은 Submetacentric chromosome, $Mdh-1^{MS}$型은 metacentric chromosome으로 큰 차이를 보였다. 따라서, $Mdh-1^{MM}$型은 6쌍의 metacentric chromosome과 6쌍의 submetacentric chromosome 및 12쌍의 acrocentric chromosome으로 구성되어 있으나, $Mdh-1^{MS}$型은 7쌍의 metacentric chromosome과 5쌍의 submetacentric chromosome 및 12쌍의 acrocentric chromosome으로 구성되어 있음을 알수 있다. 이러한 sympatric한 지역에서 이들 두型의 hybrid型이 존재하지 않는 점으로 보아 이들사이에는 生殖的 隔離가 일어난 것으로 추측된다.

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Isolation of Cold Acclimation-related Genes in Wheat Chromosome Substitution Line 5D

  • Chun, Jong Un;Jeong, In Ho
    • 한국육종학회지
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    • 제40권3호
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    • pp.234-242
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    • 2008
  • To identify low temperature-induced genes of wheat chromosome substitution line 5D, suppression subtractive hybridization (SSH) was performed with mRNAs from leaf samples that treated with low temperature ($4^{\circ}C$). A cDNA library was constructed using mRNA isolated from wheat chromosome substitution line 5D leaves treated with low temperature ($4^{\circ}C$). The nucleotide and deduced amino acid sequences of the putative gene products were compared. wfr-9 and wfr-32 showed identity over 90% related to vernalization gene. Other two genes, wfr-77 and wfr-83 which is related to freezing-resistant gene have also identity over 90%. This result suggest that those genes may be transcribed into antifreeze proteins which are accumulated within leaf apoplasts, when wheat chromosome substitution line 5D is acclimated during low temperature treatment.

Generation of FISH Probes Using Laser Microbeam Microdissection and Application to Clinical Molecular Cytogenetics

  • Shim, Sung-Han;Kyhm, Jee-Hong;Chung, Sung-Ro;Kim, Seung-Ryong;Park, Moon-Il;Lee, Chul-Hoon;Cho, Youl-Hee
    • Journal of Microbiology and Biotechnology
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    • 제17권7호
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    • pp.1079-1082
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    • 2007
  • Chromosome microdissection and the reverse FISH technique is one of the most useful methods for the identification of structurally abnormal chromosomes. In particular, the laser microbeam microdissection (LMM) method allows rapid isolation of a target chromosome or a specific region of chromosomes without damage of genetic materials and contamination. Isolated chromosomes were directly amplified by the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), and then the FISH probes labeled with spectrum green- or spectrum red-dUTP were generated by nick-translation. Whole chromosome painting (WCP) probes were successfully generated from only 5 copies of the chromosome. With this method, we produced 24 WCP probes for each human chromosome. We also tried to characterize a marker chromosome, which seemed to be originated from chromosome 11 on conventional banding technique. The marker chromosomes were isolated by the LMM method and analyzed by reverse FISH. We elucidated that the marker chromosome was originated from the short arm of chromosome 5 ($5p11{\to}pter$). A fully automated and computer-controlled LMM method is a very simple laboratory procedure, and enables rapid and precise characterization of various chromosome abnormalities.

닭에서 분리된 methicillin-resistant coagulase-negative staphylococci의 동정 및 staphylococcal cassette chromosome mec (SCCmec) type (Identification and staphylococcal cassette chromosome mec (SCCmec) type of methicillin-resistant coagulase-negative staphylococci isolated from chickens)

  • 공신국;육심용;이건택;김소연;홍영운;정윤택;이정화;김희정;황수명;장경수
    • 한국동물위생학회지
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    • 제33권3호
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    • pp.233-240
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    • 2010
  • Methicillin-resistant coagulase-negative staphylococci (MRCNS) were isolated from the respiratory sites of chickens in 4 farms and slaughter house located in Chungnam provinces. Isolation of coagulase-negative staphylococci (CNS) was positive for 61 (26.6%) of the 229 chickens tested, and isolation of MRCNS was positive for 17 (27.9%) of the isolated CNS. A total of 17 MRCNS isolates were selected and subjected to identification. Of the 17 MRCNS isolates selected, 6 were identified as Staphylococcus cohnii, 2 as S. saprophyticus, 3 as S. simulans, 3 as S. lentus, 2 as S. carnosus, and 1 as S. xylosus. The MRCNS isolates were resistant to many beta-lactam antibiotics, and some isolates were also resistant to macrolide and aminoglycoside antibiotics. The mecA gene was detected in some isolates of each MRCNS strains. The mecA-positive isolates were classified into five staphylococcal cassette chromosome mec (SCCmec). SCCmec types I to IV were detected in isolates from chickens.

Isolation and Characterization of Pseudomonas sp. KM10, a Cadmium- and Mercury-resistant, and Phenol-degrading Bacterium

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • 제8권4호
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    • pp.388-398
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    • 1998
  • A bacterium which is resistant to both mercury and cadmium, and also capable of utilizing phenol as a carbon and energy source, was isolated from the Kumho River sediments near Kangchang Bridge, Taegu, Korea. The isolate was labeled Pseudomonas sp. KM10 and characterized. The bacteria grew in 4 mM $CdCl_2$and in $70{\mu}M$ $HgCl_2$. The bacteria efficiently removed over 90% of 1 g/l phenol within 30 h. In the presence of 1.250 g/l phenol, the growth of the microorganism was slightly retarded and the microorganism could not tolerate 1.5 g/l phenol. Curing of plasmid from the bacteria was carried out to generate a plasmidless strain. Subsequent experiments localized the genes for phenol degradation in plasmid and the genes for mercury resistance and cadmium resistance on the chromosome. Dot hybridization and Southern hybridization under low stringent conditions were performed to identify the DNA homology. These results showed significant homologies between the some sequence of the chromosome of Pseudomonas sp. KM10 and merR of Shigella flexneri R 100, and between the some sequence of the chromosome of Pseudomonas sp. KM10 and cadA of Staphylococcus aureus pI258. The mechanism of cadmium resistance was efflux, similar to that of S. aureus pI258 cadA, and the mechanism of mercury resistance was volatilization, similar to that of S. flexneri R100 mer.

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Studies on Development of Resistant Strains to Antibiotics and Antituberculosis Agents(II) -Isolation of Rifampicin Resistant Mutants from Clostridium butyricum-

  • Kim, Hyung-Soo;Choi, Eung-Chil;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • 제11권3호
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    • pp.218-224
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    • 1988
  • The preparation of Clostridium butyricum is used as a normalizing agent for human intestinal flora. When the microbe is simultaneously used with rifampicin, it is inactivated by the antibiotic. To develop rifampicin-resistant mutants, rifampicin-sensitive strain Miyairi II 588 of C. butyricum was treated with nitrosoguanidine (NTG). To ensure stable resistance to rifampicin, we examined whether the resistance was plasmid-mediated or chromosome-mediated. It was found that the resistance of four mutant strains was not mediated by its inherent plasmid, but by the chromosomal mutation. These strains were examined for the susceptibility and resistance to other antituberculosis agents and antibiotics. The results showed that these mutants were resistant to the high concentration of the antituberculosis agents.

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Drosophila melanogaster complex내의 유전적 유연관계에 관한 연구 (Genetic Relationship within the melanogaster Complex of the Genus Drosophila)

  • 최영현;이원호
    • 생명과학회지
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    • 제9권1호
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    • pp.69-75
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    • 1999
  • Genus Drosophila에서 melanogaster complex내의 4종에 대한 유연관계 해석을 위하여, insemination test에 의한 premating isolation, 잡종형성 여부에 의한 postmating isolation, 형태 형성에 관한 영향 및 종 분화 관련 유전자의 특징 등을 대상으로 조사하였다. 종내 교배에서는 insemination rate는 96∼99% 정도였고 종간 교배에서는 정역 교배간에 심한 변이를 보였으며 D. melanogaster 암컷은 타종 수컷과의 교배에서 전반적 성공률이 높으며 D. sechellia 수컷은 타종 암컷과의 교배에서 비교적 높은 교배 성공률을 보였다. 종간 잡종 형성에서는 특히 simulans, mauritiana 및 sechellia 사이에서 임성이 완전한 암컷과 불임의 수컷이 형성되어 이들 3종이 더욱 근연임을 시사하고 있다. sex comb과 genital arch에 대한 잡종의 형태 형성에 관한 영향은 대부분이 polygene에 의하여 조절되고 있음을 알 수 있었다. D. melanogaster와 simulans에 있어서 분화 관련 유전자로 추정되는 잡종의 온도 감수적 생존도는 주로 simu-lans의 X 염색체상의 유전자에 의하여 조절되고 있음이 분석되었다.

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Transformation-associated recombination cloning에 의한 유전자 분리에 사용되는 target hook에 대한 GC content의 영향 (Effect of GC Content on Target Hook Required for Gene Isolation by Transformation-Associated Recombination Cloning)

  • 김중현;신영선;윤영호;장형진;김은아;김광섭;정정남;박인호;임선희
    • 미생물학회지
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    • 제39권3호
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    • pp.128-134
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    • 2003
  • Transformation-associated recombination (TAR) 클로닝법은 목적 유전자를 포함한 게놈 DNA와 그 유전자의 5' 또는 3' 말단 서열을 포함하고 있는 선형의 TAR vector를 동시에 출아효모의 spheroplast내로 co-penetration 시켜 상동부위에서 일어나는 재조합에 의해 환형의 Yeast Artification Chromosome(YAC)으로 분리되는 방법이다. 일반적으로 TAR 클로닝법에 의한 목적의 single-copy 유전자 분리 빈도는 전체 형질전환체의 0.01~1% 정도이다. 이러한 TAR 클로닝법을 개선하기 위하여 Tg.AC transgenic mouse를 모델계로 사용하여 유전자 분리에 대한 target hook 내의 GC content 가 미치는 영향을 조사하였다. 이러한 목적으로 한쪽에는 다양한 GC content(18~45%)를 지닌 transgene 특이적 hook을 포함하고 다른 한쪽은 B1 반복서열을 가지는 radial TAR vector를 사용하여 transgene 분리 빈도를 측정하였다. 그 결과 target hook의 GC content는 23% 이하의 경우, ~40%인 경우에 비해 두 배 정도 클로닝 빈도가 감소하였다. 따라서 TAR vector를 제작할 때, 유전자 분리에 이용되는 target hook의 GC content는 약 40% 일때 가장 적정한 것으로 나타났다. 또한 높은 target hook 내의 GC content(65%)위치분포에 의한 차이는 클로닝 빈도에 큰 영향을 미치지 않는 것으로 나타났다.