Expression of antigenic regions of a trypanosome variable surface glycoprotein in E. coli using Bal-31 nuclease digestion.
We have used the expression of a trypanosome variable surface glycoprotein (VSG) in E. coli to produce VSG serotype-specific antisera which have none of the cross-reacting specificities characteristic of antisera prepared against purified VSGs. This was accomplished by treating restriction fragments of VSG cDNAs with Bal-31 nuclease to facilitate expression of their open reading frames in the E. coli expression vector, pMR100 (4). The resultant VSG-beta-galactosidase fusion proteins possess various antigenic regions of the original VSG. This provides a rapid means for producing VSG-specific antisera for reagent use and has the capability of large scale production of antigen for immunological investigation.
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA / Q000032:analysis*; D004262:DNA Restriction Enzymes; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D000939:Epitopes / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics*; D006023:Glycoproteins / Q000235:genetics*; D007106:Immune Sera; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013045:Species Specificity; D014346:Trypanosoma brucei brucei / Q000276:immunology*; D014643:Variant Surface Glycoproteins, Trypanosoma; D001616:beta-Galactosidase / Q000235:genetics
D000939:Epitopes; D006023:Glycoproteins; D007106:Immune Sera; D014643:Variant Surface Glycoproteins, Trypanosoma; D004247:DNA; D004706:Endodeoxyribonucleases; C038407:exonuclease Bal 31; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase