title: Intraclonal mating in Trypanosoma brucei is associated with out-crossing.
abstract: To examine whether mating can occur within as well as between clones of Trypanosoma brucei, we transformed three T. brucei subspecies stocks with heterologous genes conferring resistance to either hygromycin or Geneticin and carried out a series of inter- and intraclone matings in all possible double drug combinations. Double drug-resistant hybrids were recovered from three of the six out-crosses, but not from any of the three intraclone matings. However, further analysis of cloned progeny trypanosomes from one of the out-crosses using RFLP markers, molecular karyotyping and RAPD (random amplification of polymorphic DNA) produced unequivocal evidence that intra- as well as interclone mating had occurred. The progeny of interclone mating were double drug-resistant and heterozygous at 9 of 13 loci examined. In contrast, the progeny of intraclone mating had no demonstrable input of genetic material from the hygromycin-resistant parent and were similar to the Geneticin-resistant parent for most markers, except for five loci which were heterozygous in the Geneticin-resistant parent but homozygous in these clones (aldolase THT1 glucose transporter, procyclin, tubulin and cDNA 23). In addition, PFGE showed considerable karyotypic rearrangements in these clones and loss of genetic material was evident from RAPD and VSG (variant surface glycoprotein) gene fingerprint analysis. We conclude that intraclone mating can occur in trypanosomes, but only during out-crossing, suggesting that meiosis and/or fusion are triggered by a diffusible factor.
authors.affiliation: Department of Genetics, University of Leicester, Leicester LE1 7RH, UK.
authors.forename: Wendy
authors.initials: W
authors.lastname: Gibson
authors.affiliation: Department of Pathology and Microbiology, University of Bristol Veterinary School, Langford, Bristol BS18 7DU, UK.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Winters
authors.affiliation: Department of Pathology and Microbiology, University of Bristol Veterinary School, Langford, Bristol BS18 7DU, UK.
authors.forename: Ginny
authors.initials: G
authors.lastname: Mizen
authors.affiliation: Department of Genetics, University of Leicester, Leicester LE1 7RH, UK.
authors.forename: Julia
authors.initials: J
authors.lastname: Kearns
authors.affiliation: Department of Clinical Veterinary Science, University of Bristol Veterinary School, Langford, Bristol BS18 7DU, UK.
authors.forename: Mick
authors.initials: M
authors.lastname: Bailey
mesh_terms: D000818:Animals; D003433:Crosses, Genetic; D015964:Gene Expression Regulation, Bacterial; D014170:Transformation, Genetic; D014346:Trypanosoma brucei brucei / Q000235:genetics*