Isolation of alpha- and beta-tubulin genes of Plasmodium falciparum using a single oligonucleotide probe.
An oligonucleotide probe (315) specific for the alpha- and beta-tubulin genes of Plasmodium falciparum was synthesized utilizing codon usage of P. falciparum determined from published gene sequences. By screening genomic and cDNA libraries with the oligonucleotide probe, alpha- and beta-tubulin clones were isolated. Positive clones were identified by partial sequencing and comparing the deduced amino acid sequence with the chicken brain alpha- and beta-tubulin amino acid sequences. The beta-tubulin gene was completely sequenced at the genomic level and partially at cDNA level. The deduced polypeptide is 445 amino acids long, shares 88% homology with chicken brain beta-tubulin, and contains two introns of 362 and 163 bp long, respectively. alpha- and beta-tubulin genes of P. falciparum are unlinked and dispersed; more than one copy of each gene may be present. Northern blot analysis of total RNA of the blood-stage parasite indicates the presence of three transcripts of alpha-tubulin (3.3 kb, 2.6 kb, 1.9 kb) and three transcripts of beta-tubulin gene (3.6 kb, 2.9 kb, 2.0 kb). The significance of these transcripts is presently unknown.
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Biochemistry Department, New York University Medical Center, New York 10016.
Godson
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003062:Codon; D005796:Genes; D007438:Introns; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010963:Plasmodium falciparum / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014404:Tubulin / Q000235:genetics*
D003062:Codon; D015345:Oligonucleotide Probes; D014404:Tubulin