Preparation of DNA libraries of Plasmodium falciparum for searching calmodulin binding protein genes, GOGAT and Pfmyo A.

Dary L. Mendoza, Moisés Wasserman, .

Keywords: plasmodium falciparum, malaria, calmodulin binding protein, glutamate synthase, myosin, cDNA

Abstract

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 genes coding 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 myosin clones--Pfmyo A, Pfmyo B and Pfmyo C--were identified. The clones coding 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 mRNA coding for this gene. Finally, the first report of a complete cDNA clone containing the P. falciparum myosin A is presented.

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  • Dary L. Mendoza Laboratorio de Bioquímica, Instituto Nacional de Salud, Bogotá, D.C. Laboratorio de Investigaciones Básicas en Bioquímica, LIBBIQ, Departamento de Química, Facultad de Ciencias, Universidad Nacional de Colombia, Bogotá, D.C.
  • Moisés Wasserman Laboratorio de Bioquímica, Instituto Nacional de Salud, Bogotá, D.C. Laboratorio de Investigaciones Básicas en Bioquímica, LIBBIQ, Departamento de Química, Facultad de Ciencias, Universidad Nacional de Colombia, Bogotá, D.C.
How to Cite
1.
Mendoza DL, Wasserman M. Preparation of DNA libraries of Plasmodium falciparum for searching calmodulin binding protein genes, GOGAT and Pfmyo A. biomedica [Internet]. 2002 Sep. 1 [cited 2024 May 18];22(3):241-52. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/1161

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Published
2002-09-01
Section
Original articles

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