Characterization of yeast artificial chromosomes from Plasmodium falciparum: Construction of a stable, representative library and cloning of telomeric DNA fragments Journal Article

  

Authors:	de Bruin, D.; Lanzer, M.; Ravetch, J. V.
Article Title:	Characterization of yeast artificial chromosomes from Plasmodium falciparum: Construction of a stable, representative library and cloning of telomeric DNA fragments
Abstract:	Molecular genetic studies of the human malaria parasite Plasmodium falciparum have been hampered in part due to difficulties in stably cloning and propagating parasite genomic DNA in bacteria. This is thought to be a result of the unusual A + T bias (>80%) in the parasite's DNA. Pulsed-field gel electrophoretic separation of P. falciparum chromosomes has shown that large chromosomal polymorphisms, resulting from the deletion of DNA from chromosome ends, frequently occur. Understanding the biological implications of this chromosomal polymorphism will require the analysis of large regions of genomic, and in particular telomeric, DNA. To overcome the limitations of cloning parasite DNA in bacteria, we have cloned genomic DNA from the P. falciparum strain FCR3 in yeast as artificial chromosomes. A pYAC4 library with an average insert size of approximately 100 kb was established and found to have a three to fourfold redundancy for single-copy genes. Unlike bacterial hosts, yeast stably maintain and propagate large tracts of parasite DNA. Long-range restriction enzyme mapping of YAC clones demonstrates that the cloned DNA is contiguous and identical to the native parasite genomic DNA. Since the telomeric ends of chromosomes are underrepresented in YAC libraries, we have enriched for these sequences by cloning P. falciparum telomeric DNA fragments (from 40 to 130 kb) as YACs by complementation in yeast. © 1992 Academic Press, Inc. All rights reserved.
Keywords:	nonhuman; telomere; animal; gene library; bacteria (microorganisms); molecular cloning; cloning, molecular; molecular sequence data; base sequence; plasmodium falciparum; chromosomes, fungal; electrophoresis, gel, pulsed-field; yeast artificial chromosome; dna library; blotting, southern; article; support, non-u.s. gov't; support, u.s. gov't, non-p.h.s.; dna, protozoan; genes, protozoan
Journal Title:	Genomics
Volume:	14
Issue:	2
ISSN:	0888-7543
Publisher:	Elsevier Inc.
Date Published:	1992-10-01
Start Page:	332
End Page:	339
Language:	English
DOI:	10.1016/s0888-7543(05)80223-x
PUBMED:	1427849
PROVIDER:	scopus
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0026468543&doi=10.1016%2fS0888-7543%2805%2980223-X&partnerID=40&md5=206835351ccd019266a6fab287afde74
Notes:	Article -- Export Date: 30 July 2019 -- Source: Scopus

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