Cloning and sequencing of a cDNA fragment from Plasmodium chabaudi chabaudi that contains repetitive sequences coding for a potentially lysine-rich aspartic acid-rich protein.
Screening of a cDNA library (prepared in lambda gt11) of the blood stages of Plasmodium chabaudi chabaudi (AS) with immune serum has revealed an antigen the elicits a strong antibody response in infected mice. The clone (clone 6) expressing that antigen contains a 0.7 kb insert and produces a beta-galactosidase fusion protein of about 150 kDa. In Western blot analysis performed on parasite extracts, monoclonal antibodies and polyclonal sera prepared against the fusion protein revealed that the fusion protein contains part of a malarial protein of 93 kDa. Northern hybridization with clone 6 insert as probe detected a plasmodial RNA of about 3.2 kb, which could well code for a protein of this size. The insert hybridized to a single EcoRI fragment and a single HindIII fragment in genomic Southern blotting, suggesting that the gene is present in one copy in the P. chabaudi genome. The DNA sequence of clone 6 insert predicts a hydrophilic, acidic polypeptide consisting of seven repeats of 23-34 amino acids rich in lysine (24%) and aspartic acid (17.5%).
Hartz
D
D
Max-Planck-Institut für Immunbiologie, Freiburg, Germany.
Ayane
M
M
Chluba-De Tapia
J
J
Wirbelauer
C
C
Langhorne
J
J
Gillard-Blass
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D001224:Aspartic Acid; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D015723:Gene Library; D016678:Genome; D008239:Lysine; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016788:Plasmodium chabaudi / Q000235:genetics*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics*; D016053:RNA, Protozoan / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012091:Repetitive Sequences, Nucleic Acid; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D011993:Recombinant Fusion Proteins; D001224:Aspartic Acid; D001616:beta-Galactosidase; D008239:Lysine