TY  - BOOK
AU  - Kordai Sowa,M.P.
AU  - Sharling,L.
AU  - Humphreys,G.
AU  - Cavanagh,D.R.
AU  - Gregory,W.F.
AU  - Fenn,K.
AU  - Creasey,A.M.
AU  - Creasey,A.M.
TI  - High throughput immuno-screening of cDNA expression libraries produced by in vitro recombination; exploring the Plasmodium falciparum proteome
KW  - PLASMODIUM FALCIPARUM
KW  - CDNA
KW  - IN VITRO RECOMBINATION
KW  - PROTEIN MACRO-ARRAY
N2  - Improved Plasmodium falciparum cDNA expression libraries were constructed by combining mRNA oligo-capping with in vitro recombination and directional cloning of cDNA inserts into a plasmid vector that expresses sequences as thioredoxin fusion proteins. A novel procedure has also been developed for the rapid identification of seropositive clones on high-density filters, using direct labelling of P. falciparum immune immunoglobulin with fluorescein isothiocynate (FITC). This approach combines the advantages of recombination-assisted cDNA cloning with high throughput, non-radioactive serological screening of expression libraries. Production of replicate colony matrices allows the identification of antigens recognised by different pools with different specificities from residents of a malaria endemic region. Analyses ofDNAsequences derived from sero-reactive colonies indicate that this is an effective method for producing recombinant proteins that react with antibodies from malaria-exposed individuals. This approach permits the systematic construction of a database of antigenic proteins recognised by sera from malaria-exposed individuals
UR  - https://drive.google.com/file/d/16z7CnuDjltYaDM9Y2rm17rgEvbwuFLUG/view?usp=drivesdk
ER  -