Isolation and identification of six Pneumocystis carinii genes utilizing codon bias.
Pneumocystis carinii pneumonia (PCP) is a leading cause of death among AIDS patients in the United States. Our analysis of P. carinii protein-coding genes has revealed a significant A + T codon bias. Polymerase chain reaction (PCR) was utilized to isolate and identify the genes encoding calmodulin, beta-tubulin, DNA polymerase II, and RNA polymerases I, II and III from P. carinii. Primer pairs were designed to incorporate P. carinii codon preference to known conserved protein regions from other organisms. This strategy should be useful for a large variety of P. carinii genes and assist in the comprehensive analysis of the genomic structure of this important pathogen.
Fletcher
L D
LD
Department of Pathology, School of Medicine, University of North Carolina, Chapel Hill 27599.
Berger
L C
LC
Peel
S A
SA
Baric
R S
RS
Tidwell
R R
RR
Dykstra
C C
CC
D000595:Amino Acid Sequence; D001483:Base Sequence; D002147:Calmodulin / Q000235:genetics; D003062:Codon / Q000032:analysis*; D004257:DNA Polymerase II / Q000235:genetics; D004271:DNA, Fungal / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005787:Gene Frequency; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011010:Pneumocystis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D014404:Tubulin / Q000235:genetics
D002147:Calmodulin; D003062:Codon; D004271:DNA, Fungal; D015345:Oligonucleotide Probes; D014404:Tubulin; D012321:DNA-Directed RNA Polymerases; D004257:DNA Polymerase II