• Title/Summary/Keyword: hybridization(FISH)

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Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • Proceedings of the Korean Society of Life Science Conference
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    • 2003.10a
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    • pp.14-27
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    • 2003
  • The genomes of Arabidopsis and rice have been fully sequenced. Genomic sequencing provides global information about genome structure and organization. A comprehensive research account of our recent studies conducted on genome painting, comparative genomics and genome fusion is provided in order to project the prospects of plant cytogenetic research in post-genomics era. Genome analysis by GISH using genome painting is demonstrated as an excellent means suitable for visualization of a whole genome, since total genomic DNA representing the overall molecular composition of the genome is used as a probe. FISH on extended DNA fibers has been developed for high-resolution FISH and has contributed to determining the copy number and order of genes. We have also mapped a number of genes involving starch synthesis on wheat chromosomes by FISH and compared the position of these genes on linkage map of rice. Macro synteny between wheat and rice can be observed by comparing the location of these genes in spite of the fact that the size of DNA per chromosome differs by 20 fold in two. Moreover, to approach our goal towards making bread and udon noodles from rice flour in future by incorporating bread making and the noodle qualifies in rice, we have been successful in introducing large genomic DNA fragments containing agronomically important genes of wheat into a rice by successive introduction of large insert BAC clones, there by expanding genetic variability in rice. We call this method genome fusion.

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Evaluation of Peptide Nucleic Acid Probe-Based Fluorescence In Situ Hybridization for the Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Clinical Respiratory Specimens (임상 객담검체에서 Peptide Nucleic Acid Probe를 이용한 결핵과 비결핵 항산균의 구분)

  • Lee, Seung Hee;Kim, Shine Young;Kim, Hyung Hoi;Lee, Eun Yup;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.18 no.2
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    • pp.37-43
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    • 2015
  • Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

Characteristics of Microbial Distribution of Nitrifiers and Nitrogen Removal in Membrane Bioreactor by Fluorescence in situ Hybridization (막/생물반응기에서 Fluorescence in situ Hybridization 기법을 이용한 질산화 미생물 분포특성 및 질소제거 연구)

  • Lim Kyoung-Jo;Kim Sun-Hee;Kim Dong-Jin;Cha Gi-Cheol;Yoo Ik-Keun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.257-264
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    • 2006
  • An aerobic submerged membrane bioreactor (MBR) treating ammonium wastewater was studied in respect of nitrification characteristics and distribution of nitrification bacteria over a period of 350 days. MBR was fed with ammonium concentration of 500-1000 mg $NH_4-N/L$ at a nitrogen load of $1-2kg\;N/m^3{\cdot}d$. Overall ammonium oxidation rate increased with dissolved oxygen (DO) concentration, temperature, and sludge retention time (SRT). Under a higher concentration of free ammonia ($NH_3-N$) due to the decrease of ammonium oxidation rate, the nitrite ratio ($NO_2-N/NO_x-N$) in the effluent increased. The sudden collapse of nitrification efficiency accompanied by sludge foaming and the increase of sludge volume index (SVI) was observed unexpectedly during the operation. At the later stage of operation, additional carbon source was fed to the MBR and resulted in twice higher value of SVI and the decrease of ammonium oxidation rate. In fluorescence in situ hybridization (FISH) analysis, genus Nitrosomonas which is specifically hybridized with probe NSM156 was initially the dominant ammonia oxidizing bacteria and the amount of Nitrosospira gradually increased. Nitrospira was the dominant nitrite oxidizing bacteria during whole operational period. Significant amount of Nitrobacter was also detected which might due to the high concentration of nitrite maintained in the reactor.

Karyotype Analysis and Physical Mapping of rDNAs Using McFISH in Jeffersonia dubia Benth (깽깽이풀의 핵형분석과 McFISH를 이용한 rDNA의 물리지도 작성)

  • Kim, Soo-Young;Choi, Hae-Woon;Koo, Dal-Hoe;Kim, Chan-Soo;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.48-51
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    • 2005
  • Karyotype analysis and chromosomal locailization of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) were carried out in Jeffersonia dubia Benth., which is one of medicinal plants belonging to Berberidaceae. The somatic metaphase chromosome number was 2n=2x=12 and the size of chromosomes ranged $1.95{\sim}3.50\;{\mu}m$. The chromosome complement consisted of two pairs of metacentrics (chromosomes 1 and 3), two pairs of submetacentrics (chromosomes 2 and 4) and two pairs of subtelocentrics (chromosomes 5 and 6). In McFISH, one pair of 45S rDNA site was detected on the centromeric region of chromosome 2 and three pairs of 5S rDNA sites were detected on the short arm of chromosomes 4, 5 and 6, respectively.

Interspecific hybridization in seahorses: artificially produced hybrid offspring of Hippocampus kuda and Hippocampus reidi

  • Han, Sang-Yun;Rho, Sum;Noh, Gyeong Eon;Kim, Jin-Koo
    • Fisheries and Aquatic Sciences
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    • v.21 no.5
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    • pp.11.1-11.8
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    • 2018
  • Interspecific hybridization experiments were conducted between the common seahorse Hippocampus kuda (male) and the slender seahorse H. reidi (female) during artificial rearing to develop a new aquarium fish with unique polyandrous mating. Molecular analysis via mitochondrial DNA (mtDNA) cytochrome b and nuclear DNA (ncDNA) ribosomal protein S7 gene supported the hybridization between the two species, and the hybrid also showed morphological characteristics of both species. Juveniles of H. kuda have dense melanophores on the whole body or only on the trunk and tail, whereas juveniles of H. reidi have thin melanophores on the whole body or present in stripes only along their prominent trunk and tail rings. However, all the hybrid juveniles had dense melanophores only on the tail, with the striped trunk rings, thus showing an intermediate pattern, and these patterns were limited to the fairly early stage of development (1-10 days old). In contrast, the two eye spines in the hybrid were apparent after 9 days old, which were not inherited from H. kuda (one eye spine), but from H. reidi (two eye spines). According to LOESS (local regression) analysis, the growth rate increased between 20 and 25 days, and the hybrids grew faster than H. kuda when they entered the explosive second phase of growth between 25 and 45 days for all the seahorses. This study highlights the hybridization between H. kuda and H. reidi may contribute to the improved taxonomic information of young seahorses.

Chromosomal Localization and Distribution of the Telomeric DNA in Cattle and Pigs (소, 돼지 염색체의 telomeric DNA 분포 양상)

  • Sohn, S.H.;Multani, A.S.;Pathak, S.;Cho, E.J.;Ha, H.B.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.547-554
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    • 2004
  • Telomeres are nucleoprotein structures at the ends of chromosomes consisting of tandem repeat sequences of . (TTAGGG)n. Telomeres serve as guardians of the genome, protect individual chromosomes within the nucleus, and help in meiotic pairing of homologous chromosomes. To investigate the telomere distributions of cattle and pig chromosomes, fluorescence in situ hybridization(FISH) was carried out on metaphase spreads of in vitro fibroblast cultures from Holstein and Landrace using a human telomeric DNA repeat probe. Results indicate that the distinct double spots on both ends of chromosomes of cattle and pigs were observed. In cattle, there was a random variation in the intensity of telomere signals among chromosomes. In pigs, an interstitial telomeric signal was observed on the chromosome 6q1 of all the cells examined. According to quantitative fluorescence in situ hybridization(Q-FISH) analysis, some chromosomes had consistently much more telorneres at one end of chromosomes. In general, both species had consistently much more telomeres at q-end than p-end on most of chromosomes. The relative amount of telomeres on bovine chromosomes was higher than that on pig chromosomes. In additions, Y chromosome had the highest relative amount of telorneres in cattle and pigs.

Current Status of Comparative Mapping in Livestock

  • Lee, J.H.;Moran, C.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1411-1420
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    • 2003
  • Comparative maps, representing chromosomal locations of homologous genes in different species, are useful sources of information for identifying candidate disease genes and genes determining complex traits. They facilitate gene mapping and linkage prediction in other species, and provide information on genome organization and evolution. Here, the current gene mapping and comparative mapping status of the major livestock species are presented. Two techniques were widely used in comparative mapping: FISH (Fluorescence In Situ Hybridization) and PCR-based mapping using somatic cell hybrid (SCH) or radiation hybrid (RH) panels. New techniques, using, for example, ESTs (Expressed Sequence Tags) or CASTS (Comparatively Anchored Sequence Tagged Sites), also have been developed as useful tools for analyzing comparative genome organization in livestock species, further enabling accurate transfer of valuable information from one species to another.

A New Acheilognathine Fish, Acheilognathus somjinensis (Pisces: Cyprinidae) from Korea (한국산 납자로속(잉어과) 어류 1 신종)

  • 김익수;김치홍
    • Animal Systematics, Evolution and Diversity
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    • v.7 no.2
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    • pp.189-194
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    • 1991
  • A new speices of cyprinid fish, Acheilognathus somjinensis is described based on the specimens collected in the Somjin River, Sinpyong-myon, Imsil -gun , Chollabuk-do province of Korea. In general morphology it is similar to A. koreensis. however, the new speices differs from the latter in having longer ovipositor of female in spawing season and showing pear-shaped eggs. In hybridization experiments, it was found that A.somjinensis is repoductively isolated from A.koreensis.

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Early Life History Characteristics of an Induced Hybrid Between Rhodeus ocellatus and Rhodeus notatus (흰줄납줄개(Rhodeus ocellatus)와 떡납줄갱이(Rhodeus notatus) 잡종의 초기생활사 특징)

  • Jae-Min Park;Kyeong-Ho Han
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.4
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    • pp.549-557
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    • 2023
  • This study aimed to investigate taxonomic differences in the early life history characteristics of Rhodeus ocellatus and Rhodeus notatus through an interspecific hybridization experiment. The hatching rates of the cross-bred eggs were 50% for the ON cross (R. ocellatus ♀×R. notatus♂) and 67% for the NO cross (R. notatus♀×R. ocellatus♂). Interestingly, the hatching time of the hybrids was longer than that of their parents. The newly hatched larvae exhibited distinct differences in their physical characteristics. The head of the adult fish protruded forward more than that of the ON cross. The pterygoid process was 45° for the ON cross and 90° for the NO cross. Furthermore, the tail shape of the ON and NO crosses closely resembled that of R. notatus.