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Two-dimensional screening of the Wageningen chicken BAC library   总被引:10,自引:0,他引:10  
We have constructed a Bacterial Artificial Chromosome (BAC) library that provides 5.5-fold redundant coverage of the chicken genome. The library was made by cloning partial HindIII-digested high-molecular-weight (HMW) DNA of a female White Leghorn chicken into the HindIII site of the vector pECBAC1. Several modifications of standard protocols were necessary to clone efficiently large partial HindIII DNA fragments. The library consists of 49,920 clones arranged in 130 384-well plates. An average insert size of 134 kb was estimated from the analysis of 152 randomly selected BAC clones. The average number of NotI restriction sites per clone was 0.77. After individual growth, DNA was isolated of the pooled clones of each 384-well plate, and subsequently DNA of each plate was isolated from the individual row and column pools. Screening of the Wageningen chicken BAC library was performed by two-dimensional PCR with 125 microsatellite markers. For 124 markers at least one BAC clone was obtained. FISH experiments of 108 BAC clones revealed chimerism in less than 1%. The number of different BAC clones per marker present in the BAC library was examined for 35 markers which resulted in a total of 167 different BAC clones. Per marker the number of BAC clones varied from 1 to 11, with an average of 4.77. The chicken BAC library constitutes an invaluable tool for positional cloning and for comparative mapping studies. Received: 26 October 1999 / Accepted: 6 January 2000  相似文献   

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Roots in the Wageningen Rhizolab are observed using two methods: (i) non-destructively, using horizontal, glass minirhizotrons at intervals of 14 days between observations; (ii) with destructive sampling using augers on three dates in the season. This paper reports changes with depth and time in root numbers per unit interface area of the minirhizotron tube (number of intersections) of four crop species (wheat, Brussels sprouts, leek and potato). The number of root intersections of Brussels sprouts, wheat and potato declined with depth at any time, whereas leek showed a different pattern because maximum root growth was observed at a depth of 10–20 cm. Root density generally decreased in the following order: Brussels sprouts, wheat, potato and leek. Plots of root length densities, Lrv(cm. cm-3), obtained by auger sampling, versus the number of intersections showed considerable variation in slope with species, time in the season and year, implying that a single, universal equation to convert minirhizotron observations into volumetric root densities does not exist. Causes of variation in the slopes are discussed. It is concluded that limited auger sampling combined with minirhizotron observations yield adequate quantitative estimates of relevant root properties.  相似文献   

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During the second half of the 19th century Julius von Sachs established the main principles of the photosynthetic production of sugars. From then, a growing number of biochemists and physiologists attended to the process, that appeared like a "black box", in order to detect what came in and what went out of it. The English group of Frederick Blackman gave a remarkable contribution in individuating the close connection between temperature, light and CO2 concentration. Later, the great importance of light was stressed by Otto Warburg, who evaluated the radiant energy necessary to the process in terms of quantum theory. The biochemical mechanism of photosynthesis was interpreted by the main European schools on the basis of Adolf Baeyer's suggestion which posed formaldehyde as the core of the process. Formaldehyde's theory hold engaged the biochemists for about fifty years although some voices rose up against it. However, nobody could put forward more coherent theories until the 1940s, when Sam Ruben and Martin Kamen individuated the cyclic pattern of the process. Ultimately, the first thirty years of the 20th century must be seen as a preliminary stage studded with light and shade even if, in spite of controversial trends, several findings of remarkable interest became to disclose that "black box" as we know today chlorophyll photosynthesis.  相似文献   

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