Plate-based transfection and culturing technique for genetic manipulation of Plasmodium falciparum

被引：17

作者：

Caro, Florence ^{[1
]}

Miller, Mathew G. ^{[2
]}

DeRisi, Joseph L. ^{[1
,3
]}

机构：

[1] Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94143 USA

[2] Codexis Inc, Redwood City, CA 94063 USA

[3] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA USA

来源：

MALARIA JOURNAL | 2012年 / 11卷

关键词：

malaria; Plasmodium falciparum; transfection; 96-well plate; SACCHAROMYCES-CEREVISIAE GENOME; DOUBLE CROSSOVER RECOMBINATION; NEGATIVE SELECTION; HUMAN ERYTHROCYTES; MALARIA PARASITES; EXPRESSION; DELETION; TRANSFORMATION; INTEGRATION; PROTEINS;

D O I：

10.1186/1475-2875-11-22

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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.

引用

页数：10

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