Preparación de genotecas de ADNc de Plasmodium falciparum para la búsqueda de los genes de proteínas de unión a calmodulina, GOGAT y Pfmyo A / Preparation of DNA libraries of Plasmodium falciparum for searching calmodulin binding protein genes, GOGAT and Pfmyo A
A cDNA library of Plasmodium falciparum (Colombian strain FCB2) asexual stage was constructed in the lambda ZipLox vector. The lambda ZipLox library and a lambda ZAPII (Dd2 strain) were screened for genescoding for proteins that bind with or are related to calmodulin (CaM). Screening was accomplished with Hot start PCR assays and hybridization with radiolabeled probes. Actin I, CaM, glutamate synthase (GOGAT) and the three myosinclones--Pfmyo A, Pfmyo B and Pfmyo C--were identified. The clonescoding for actin I, CaM and GOGAT were retrieved from the lambda ZipLox library, and the GOGAT and Pfmyo A clones from the lambda ZAP II library. The GOGAT clone contained an insert of 2,413 base pairs corresponding to 24.8 of the reported sequence. The Pfmyo A insert was 2,457 base pairs long, and represented the complete mRNAcoding for this gene. Finally, the first report of a complete cDNAclone containing the P. falciparum myosin A is presented.