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PMID: 2201894 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of the URA5 gene from Cryptococcus neoformans var. neoformans and its use as a selective marker for transformation.

Molecular and cellular biology ·Vol. 10 ·No. 9 ·1990-09-00 ·Pages 4538-44

Edman JC, Kwon-Chung KJ

Abstract

A cDNA encoding Cryptococcus neoformans orotidine monophosphate pyrophosphorylase (OMPPase) has been isolated by complementation of the cognate Escherichia coli pyrE mutant. The cDNA was used as a probe to isolate a genomic DNA fragment encoding the OMPPase gene (URA5). By using electroporation for the introduction of plasmid DNA containing the URA5 gene, C. neoformans ura5 mutants could be transformed at low efficiency. Ura+ transformants obtained with supercoiled plasmids containing the URA5 gene showed marked mitotic instability and contained extrachromosomal URA5 sequences, suggesting limited ability to replicate within C. neoformans. Transformants obtained with linear DNA were of two classes: stable transformants with integrated URA5 sequences, and unstable transformants with extrachromosomal URA5 sequences.

MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Southern Cryptococcus/genetics Cryptococcus neoformans/enzymology,genetics DNA, Fungal/genetics Escherichia coli/genetics Gene Library Genes, Fungal Molecular Sequence Data Orotate Phosphoribosyltransferase/genetics Restriction Mapping Transformation, Genetic
Chemicals
DNA, Fungal Orotate Phosphoribosyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Edman J C
Hormone Research Institute, University of California, San Francisco 94143-0534.
Kwon-Chung K J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.9.4538-4544.1990
Published
1990-09-00
Pages
4538-44
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361041
Subset
IM
Databases
GENBANK
M34606
External Links
PubMed source DOI: 10.1128/mcb.10.9.4538-4544.1990 PMC Full Text View journal details
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