Specific cloning of human DNA as yeast artificial chromosomes by transformation-associated recombination.
نویسندگان
چکیده
منابع مشابه
A new vector for recombination-based cloning of large DNA fragments from yeast artificial chromosomes.
The functional analysis of genes frequently requires manipulation of large genomic regions embedded in yeast artificial chromosomes (YACs). We have designed a yeast-bacteria shuttle vector, pClasper, that can be used to clone specific regions of interest from YACs by homologous recombination. The important feature of pClasper is the presence of the mini-F factor replicon. This leads to a signif...
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Large regions of human DNA can be cloned and mapped in yeast artificial chromosomes (YACs). Overlapping YAC clones can be used in order to reconstruct genomic segments in vivo by meiotic recombination. This is of importance for reconstruction of a long gene or a gene complex. In this work we have taken advantage of yeast protoplast fusion to generate isosexual diploids followed by mitotic cross...
متن کاملIntegrative selection of human chromosome-specific yeast artificial chromosomes.
Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of...
متن کاملConstruction of human chromosome 21-specific yeast artificial chromosomes.
Chromosome 21-specific yeast artificial chromosomes (YACs) have been constructed by a method that performs all steps in agarose, allowing size selection by pulsed-field gel electrophoresis and the use of nanogram to microgram quantities of DNA. The DNA sources used were hybrid cell line WAV-17, containing chromosome 21 as the only human chromosome and flow-sorted chromosome 21. The transformati...
متن کاملMeiotic Recombination on Artificial Chromosomes in Yeast Lyle
We have examined the meiotic recombination characteristics of artificial chromosomes in Saccharomyces cereuisiae. Our experiments were carried out using minichromosome derivatives of yeast chromosome ZZZ and yeast artificial chromosomes composed primarily of bacteriophage X DNA. Tetrad analysis revealed that the artificial chromosomes exhibit very low levels of meiotic recombination. However, w...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1996
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.93.1.491