Expression of single-domain soluble and disulfide-folded PfEMP1 antigens in the Escherichia coli SHuffle expression system
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Expression of single-domain soluble and disulfide-folded PfEMP1 antigens in the Escherichia coli SHuffle expression system. / Quintana, Maria Del Pilar.
Malaria Immunology: Targeting the Surface of Infected Erythrocytes. Vol. 2470 Humana Press, 2022. p. 273-282 (Methods in molecular biology (Clifton, N.J.)).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Expression of single-domain soluble and disulfide-folded PfEMP1 antigens in the Escherichia coli SHuffle expression system
AU - Quintana, Maria Del Pilar
N1 - © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - The genome of Plasmodium falciparum has an A/T content of around 81%. This, together with a high cysteine content and the high molecular weight of several proteins, make the expression of recombinant parasite proteins in heterologous systems challenging. P. falciparum erythrocyte membrane protein 1 (PfEMP1) is a family of proteins composed of several Duffy-binding like (DBL) and cysteine-rich inter-domain region (CIDR) domains involved in cytoadhesion to human host receptors and development of severe malaria. Expression of correctly folded single- and multiple-domain PfEMP1 fragment regions containing cysteines forming disulfide bonds, remains particularly difficult. Nevertheless, expression of single DBL and CIDR domains has been successful and this protocol describes the expression and purification of single-domain soluble PfEMP1 fragments using the Escherichia coli SHuffle expression system.
AB - The genome of Plasmodium falciparum has an A/T content of around 81%. This, together with a high cysteine content and the high molecular weight of several proteins, make the expression of recombinant parasite proteins in heterologous systems challenging. P. falciparum erythrocyte membrane protein 1 (PfEMP1) is a family of proteins composed of several Duffy-binding like (DBL) and cysteine-rich inter-domain region (CIDR) domains involved in cytoadhesion to human host receptors and development of severe malaria. Expression of correctly folded single- and multiple-domain PfEMP1 fragment regions containing cysteines forming disulfide bonds, remains particularly difficult. Nevertheless, expression of single DBL and CIDR domains has been successful and this protocol describes the expression and purification of single-domain soluble PfEMP1 fragments using the Escherichia coli SHuffle expression system.
KW - Antigens, Protozoan
KW - Cysteine/metabolism
KW - Disulfides/metabolism
KW - Erythrocytes/metabolism
KW - Escherichia coli/genetics
KW - Humans
KW - Malaria, Falciparum
KW - Plasmodium falciparum/metabolism
KW - Protozoan Proteins/metabolism
KW - Recombinant Proteins/genetics
U2 - 10.1007/978-1-0716-2189-9_20
DO - 10.1007/978-1-0716-2189-9_20
M3 - Book chapter
C2 - 35881352
SN - 978-1-0716-2188-2
VL - 2470
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 273
EP - 282
BT - Malaria Immunology
PB - Humana Press
ER -
ID: 327471856