Schistosoma mansoni: cloning of antigen gene sequences in Escherichia coli.
Fischer rat protective antiserum (F-2x) prepared from Schistosoma mansoni-infected rats was used to screen an adult worm cDNA library constructed in a lambda gt11 bacteriophage expression vector. This led to the isolation of several clones yielding proteins reactive with antibodies in the infection serum. Counter-screening of these clones with Wistar-Furth rat nonprotective antiserum (W-2x) enabled identification of clones either uniquely or preferentially reacting with F-2x, in addition to clones of nearly equal reactivity with both antisera. Six clones were further characterized. Five expressed beta-galactosidase/S. mansoni fusion proteins which migrated more slowly in sodium dodecyl sulfate-polyacrylamide gel electrophoresis than beta-galactosidase and all were reactive in a Western immunoblot assay. The cDNA insert sizes in the clones ranged from 150 to 900 base pairs. Rabbit antibodies prepared against fusion proteins from three of the clones recognized biosynthetically radio-labeled 4-week worm proteins of sizes 20, 38, and 70 kDa, respectively. The 20- and 38-kDa proteins were among the protein antigens uniquely recognized by the F-2x protective antiserum. These proteins are therefore candidates for protective vaccine antigens and the recombinant lambda clones are now serving as useful reagents for obtaining the corresponding nucleotide gene sequences.
Division of Biology and Medicine, Brown University, Providence, Rhode Island 02912.
M
M
el-Sherbeini
K A
KA
Bostian
P M
PM
Knopf
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics*; D001345:Autoradiography; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D015723:Gene Library; D007106:Immune Sera / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D051381:Rats; D011916:Rats, Inbred F344; D011920:Rats, Inbred WF; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D015183:Restriction Mapping; D012550:Schistosoma mansoni / Q000235:genetics* / Q000276:immunology
D000947:Antigens, Helminth; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D004247:DNA