Journal article
Construction of a human chromosome 4 YAC pool and analysis of artificial chromosome stability
Nucleic acids research, Vol.20(13), pp.3419-3425
1992
DOI: 10.1093/nar/20.13.3419
PMCID: PMC312498
PMID: 1630913
Abstract
In order to construct a human chromosome 4-specific YAC library, we have utilized pYAC4 and a mouse/human hybrid cell line HA(4)A in which the only human chromosome present is chromosome 4. From this cell line, approximately 8Mb of chromosome 4 have been cloned. The library includes 65 human-specific clones that range in size from 30kb to 290kb, the average size being 108kb. In order to optimize the manipulation of YAC libraries, we have begun to investigate the stability of YACs containing human DNA in yeast cells; these studies will also determine if there are intrinsic differences in the properties of chromosomes containing higher eukaryotic DNAs. We are examining two kinds of stability: 1] mitotic stability, the ability of the YAC to replicate and segregate properly during mitosis, and 2] structural stability, the tendency of the YAC to rearrange. We have found that the majority of YACs examined are one to two orders of magnitude less stable than authentic yeast chromosomes. Interestingly, the largest YAC analyzed displayed a loss rate typical for natural yeast chromosomes. Our results also suggest that increasing the length of an artificial chromosome improves its mitotic stability. One YAC that showed a very high frequency of rearrangement by mitotic recombination proved to be a mouse/human chimera. In contrast to studies using total human DNA, the frequency of chimeras (i.e., mouse/human) in the YAC pool appeared to be low.
Details
- Title: Subtitle
- Construction of a human chromosome 4 YAC pool and analysis of artificial chromosome stability
- Creators
- Heidi Major SleisterKathleen A MillsSue E BlackwellAnn M KillaryJeffrey C MurrayRobert E Malone
- Resource Type
- Journal article
- Publication Details
- Nucleic acids research, Vol.20(13), pp.3419-3425
- DOI
- 10.1093/nar/20.13.3419
- PMID
- 1630913
- PMCID
- PMC312498
- NLM abbreviation
- Nucleic Acids Res
- ISSN
- 0305-1048
- eISSN
- 1362-4962
- Language
- English
- Date published
- 1992
- Academic Unit
- Anatomy and Cell Biology; Stead Family Department of Pediatrics; Epidemiology; Biology; Pediatric Dentistry; Craniofacial Anomalies Research Center; Dental Research; Holden Comprehensive Cancer Center
- Record Identifier
- 9984025676602771
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