Characterisation and translation studies of messenger RNA from the human malaria parasite Plasmodium falciparum and construction of a cDNA library

John E. Hyde, Michael Goman, Roger Hall, Arve Osland, Ian A. Hope, Gordon Langsley, J. Werner Zolg, John G. Scaife

    Research output: Contribution to journalArticlepeer-review

    Abstract

    RNA was isolated from trophozoites, schizonts and mixed populations of Plasmodium falciparum. 5% of the total was poly(A+) message, of average length 1.2 kb (10-12 kb maximum) and a poly(A) content of 10%. The mRNA fractions could be translated in vitro by reticulocyte lysates supplemented either with exogenous or P. falciparum tRNA. The patterns from two independent isolates, one cloned (T9-94) and one uncloned (K1) were virtually identical. Major translation products from 16-230 kDa have been measured. The most abundant is lactate dehydrogenase (34.8 kDa). Trophozoite mRNA codes principally for proteins of ≤93 kDa, while schizont mRNA codes for additional proteins of higher molecular mass. There are marked similarities between the in vitro translation products and proteins synthesised in vivo in synchronous cultures. A number of schizont mRNA translation products (principally those of 230, 203, 185, 170, 115, 101 and 71 kDa) are specifically precipitated without post-translational modification by sera from humans exposed to malaria. A cDNA library has been constructed in phage λ from total poly(A+) RNA and partially characterised. About 10% of the clones derive from abundant mRNA sequences. Putative actin clones have been isolated from this library and the parasite actin mRNA sized at approx. 2.8 kb. © 1984.
    Original languageEnglish
    Pages (from-to)269-285
    Number of pages16
    JournalMolecular and biochemical parasitology
    Volume10
    Issue number3
    DOIs
    Publication statusPublished - Mar 1984

    Keywords

    • Actin clones
    • cDNA library
    • Immunoprecipitation
    • Plasmodium falciparum
    • Stage-specific mRNA
    • Translation in vitro

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