New tools to identify var sequence tags and clone full-length genes using type-specific primers to Duffy binding-like domains.

Abstract:

:Cytoadherence of infected erythrocytes is a hallmark of Plasmodium falciparum infection and a key determinant in the particular virulence of this species. Infected erythrocytes bind a variety of host receptors but certain adhesion traits are associated with more severe disease. A large, diverse protein family named P. falciparum erythrocyte membrane protein 1 (PfEMP1) is responsible for sequestration of mature stage infected erythrocytes and orchestrates parasite binding tropism. To better understand the molecular basis for malaria disease, more study is needed to identify the subset of PfEMP1 variants that contribute to basic disease phenotypes. PfEMP1 proteins have multiple receptor-like domains that group into different homology types based upon sequence similarity. Universal primers have been developed that recognize some, but not all PfEMP1 adhesion domain types. In this study, we designed and validated a new series of type-discriminatory primers to the DBL-beta, -gamma, and -delta adhesion types for epidemiological profiling. In addition, we used new primers to the var upstream region and exon 2 to demonstrate how the strategic placement of primers throughout the gene structure can be exploited to efficiently clone the var gene coding region. These new approaches provide valuable tools to gain novel insights into cytoadherence and malaria pathogenesis.

journal_name

Mol Biochem Parasitol

authors

Kraemer SM,Gupta L,Smith JD

doi

10.1016/s0166-6851(03)00099-9

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

91-102

issue

1

eissn

0166-6851

issn

1872-9428

pii

S0166685103000999

journal_volume

129

pub_type

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