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Mtambo J Van Bortel W Madder M Roelants P Backeljau T 《Experimental & applied acarology》2006,38(2-3):189-199
Five differently preserved groups of adult Rhipicephalus appendiculatus specimens were compared for quality of DNA extracted. Three methods were used to extract DNA from specimens i.e. two simple
mosquito validated DNA extraction methods and a tick validated method. Extraction of DNA from tick legs was attempted. The
quality of DNA extracted was evaluated by the success of PCR amplification of the ITS2 gene and the mitochondrial COI gene
fragment. Fresh specimens (i.e. killed just before extraction) had the highest success of DNA amplification followed by specimens
killed in ethanol and subsequently stored in the refrigerator (4 °C). There was no significant difference in amplification
success between cryopreserved and 70% ethanol preserved specimens. It was possible to amplify DNA from legs of ticks. Sequenced
ITS2 amplicon of template obtained from legs of ticks was as legible as those from whole tick extract. The two mosquito validated
DNA extraction methods showed a significantly lower amplification success than the tick validated protocol. 相似文献
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The overlapping fragments of the chromosomal DNA from black widow spider Latrodectus mactans carrying genes for high-molecular-mass protein neurotoxins, - and -latroinsectotoxins (-LIT and -LIT) and -latrotoxin (-LTX), were PCR-amplified and cloned. Restriction analysis of the PCR products showed that the distribution and sizes of the restriction fragments coincided with those deduced from the earlier sequencing of cDNAs of the corresponding genes. It thus followed that the -LIT and -LIT genes are intronless. Along with our data on the structure of the -latrocrustotoxin (-LCT), this implies that the lack of introns is a common feature of the black widow spider genes encoding high molecular mass neurotoxins. 相似文献
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Construction of a bacterial artificial chromosome library of Medicago truncatula and identification of clones containing ethylene-response genes 总被引:6,自引:0,他引:6
Y.-W. Nam R. V. Penmetsa G. Endre P. Uribe D. Kim D. R. Cook 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(3-4):638-646
To facilitate genome analysis and map-based cloning of symbiotic genes in the model legume Medicago truncatula, a bacterial artificial chromosome (BAC) library was constructed. The library consists of 30 720 clones with an average insert
size of approximately 100 kb, representing approximately five haploid-genome equivalents. The frequency of BAC clones carrying
inserts of chloroplast DNA was estimated to be 1.4%. Screening of the library with single- or low-copy genes as hybridization
probes resulted in the detection of 1–12 clones per gene. Hybridization of the library with repeated sequences such as rDNA
genes and transposon-like elements of M. truncatula revealed the presence of 60 and 374 BAC clones containing the two sequences, respectively. The BAC library was pooled for
screening by polymerase chain reaction (PCR)-amplification. To demonstrate the utility of this system, we used primers designed
from a conserved region of the ein3-like loci of Arabidopsis thaliana and isolated six unique BAC clones from the library. DNA gel-blot and sequence analyses showed that these ein3-like clones could be grouped into three classes, an observation consistent with the presence of multiple ein3-like loci in M. truncatula. These results indicate that the BAC library represents a central resource for the map-based cloning and physical mapping
in M. truncatula and other legumes.
Received: 27 July 1998 / Accepted: 5 August 1998 相似文献
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