Molecular cloning and characterization of a Leishmania donovani alpha-tubulin gene.
We have isolated a cDNA for an alpha-tubulin mRNA from L. donovani promastigotes and determined its complete nucleotide sequence. Both nucleotide and deduced amino acid sequence analysis of this cDNA showed significant similarity with a previously reported, partial sequence of an L. enriettii alpha-tubulin and the complete sequence of human alpha-tubulin. Further, the in vitro translated L. donovani alpha-tubulin gene product was specifically immunoprecipitated with a monoclonal antibody against human alpha-tubulin. Northern blot analysis revealed that there was little change in the expression of the L. donovani alpha-tubulin RNA during parasite differentiation from promastigote to the in vitro grown "amastigote" form. Southern blot analysis revealed a simple genomic organization for the L. donovani alpha-tubulin gene with more than one copy of the alpha-tubulin gene in the parasite genome. To our knowledge, this is the first complete sequence of an alpha-tubulin for Leishmania to be reported in the literature.
Joshi
M
M
Laboratory of Molecular Pharmacology, Division of Hematologic Products, CBER, US Food and Drug Administration, Bethesda, Maryland, USA.
Dwyer
D M
DM
Nakhasi
H L
HL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018628:Gene Dosage; D018507:Gene Expression Regulation, Developmental; D017125:Genes, Protozoan / Q000235:genetics*; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D016053:RNA, Protozoan / Q000032:analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014404:Tubulin / Q000096:biosynthesis / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D016053:RNA, Protozoan; D014404:Tubulin