title: HygR and PurR plasmid vectors for episomal transfection of Trypanosoma cruzi.
abstract: This work describes the development and functional testing of two episomes for stable transfection of Trypanosoma cruzi. pHygD contained the 5'- and 3'- flanking regions of the gene encoding the cathepsin B-like protease of T. cruzi as functional trans-splicing and polyadenylation signals for the hygR ORF. Evidence is presented to support extrachromosomal maintenance and organization as tandem repeats in transfected parasites. pPac was derived from pHygD by replacement of the entire hygR ORF with a purR coding region. The ability to modify pHygD and the availability of the complete DNA sequence make these plasmids useful tools for the genetic manipulation of T. cruzi.
authors.affiliation: Laboratório Multidisciplinar de Pesquisa em Doença de Chagas, Faculdade de Medicina, Universidade de Brasília, Brasília, DF, Brazil.
authors.forename: Otávio T
authors.initials: OT
authors.lastname: Nóbrega
authors.forename: Jaime M
authors.initials: JM
authors.lastname: Santana
authors.forename: Nancy R
authors.initials: NR
authors.lastname: Sturm
authors.forename: Antônio R L
authors.initials: AR
authors.lastname: Teixeira
authors.forename: David A
authors.initials: DA
authors.lastname: Campbell
mesh_terms: D000818:Animals; D002934:Cinnamates; D005822:Genetic Vectors / Q000235:genetics*; D006921:Hygromycin B / Q000031:analogs & derivatives*; D008969:Molecular Sequence Data; D011691:Puromycin; D014162:Transfection / Q000379:methods*; D014349:Trypanosoma cruzi / Q000235:genetics*
chemical_list: D002934:Cinnamates; D006921:Hygromycin B; C026273:hygromycin A; D011691:Puromycin