Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum.
Malar J
; 11: 22, 2012 Jan 18.
Article
em En
| MEDLINE
| ID: mdl-22257490
ABSTRACT
Genetic manipulation of malaria parasites remains an inefficient, time-consuming and resource-intensive process. Presented here is a set of methods for 96-well plate-based transfection and culture that improve the efficiency of genetic manipulation of Plasmodium falciparum. Compared to standard protocols plate-based transfection requires 20-fold less DNA, transient transfection efficiency achieved is approximately seven-fold higher, whilst stable transfection success rate is above 90%. Furthermore the utility of this set of protocols to generate a knockout of the PfRH3 pseudogene, screened by whole-cell PCR, is demonstrated. The methods and tools presented here will facilitate genome-scale genetic manipulation of P. falciparum.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Plasmodium falciparum
/
Transfecção
/
Genética Microbiana
/
Biologia Molecular
Limite:
Humans
Idioma:
En
Revista:
Malar J
Assunto da revista:
MEDICINA TROPICAL
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos