Stage and strain specific expression of the tandemly repeated 90 kDa surface antigen gene family in Trypanosoma cruzi.
A recombinant cDNA library constructed in the expression vector lambda gtll using mRNA from the trypomastigote stage of Trypanosoma cruzi was screened with two monoclonal antibodies that have been shown to react with a 105 kDa and a 90 kDa surface antigen in trypomastigotes of the Peru and Y strains of T. cruzi. One recombinant lambda phage, designated Tcc-20, was reactive to both monoclonals. The beta-galactosidase/T. cruzi hybrid protein encoded in Tcc-20 is recognized by the monoclonal antibodies and by serum antibodies from mice infected with strains of T. cruzi which contain the 90 kDa antigen. Antibodies immunoselected from serum of mice infected with the Peru strain by adsorption to Tcc-20 fusion protein react specifically with a 90 kDa polypeptide in trypomastigote but not epimastigote lysates of T. cruzi. The mRNA complementary to the DNA insert in Tcc-20 is present only in those stages and strains of T. cruzi which express the 90 kDa surface antigen. These characteristics are strong evidence that the T. cruzi DNA fragment cloned into Tcc-20 encodes a portion of the 90 kDa surface antigen. The gene(s) which encodes this polypeptide is shown to be present in approximately 20 copies per haploid genome and most, and possibly all, of the copies are found in a tandemly linked multigene family.
Beard
C A
CA
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Wrightsman
R A
RA
Manning
J E
JE
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D005786:Gene Expression Regulation; D005796:Genes; D007118:Immunoassay; D051379:Mice; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000276:immunology; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity; D014349:Trypanosoma cruzi / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI