Open AccessCharacterisation of a novel interspersed Toxoplasma gondii DNA repeat with potential uses for PCR diagnosis and PCR‐RFLP analysis
Author(s) -
Echeverria Pablo C,
Rojas Paola A,
Martin Valentina,
Guarnera Eduardo A,
Pszenny Viviana,
Angel Sergio O
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb08984.x
Subject(s) - biology , microbiology and biotechnology , toxoplasma gondii , restriction fragment length polymorphism , genetics , pseudogene , restriction enzyme , polymerase chain reaction , dna , sequence analysis , genome , gene , antibody
A novel Toxoplasma gondii interspersed repeat element (TgIRE), present in most of the tachyzoite chromosomes, was characterised. Two regions on the TgIRE sequence showed high identity to two different T. gondii expressed sequence tag cDNAs of unknown function, which seems to be TgIRE pseudogenes. Two set of primers were designed, 2‐2′ and 2‐3, that amplify products of 1.02 and 0.62 kb, respectively. T. gondii DNA from RH and Me49 strains was amplified with TgIRE 2‐2′ primers, and the respective 1.02 kb products were digested with several endonucleases. Different fragment patterns by gel electrophoresis were found only with Mbo I. Sensitivity analysis revealed that the set 2‐3 was more sensitive than 2‐2′, detecting by gel visualisation the amount of DNA equivalent to 1 and 10 parasites, respectively.