Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-20T07:27:23.914Z Has data issue: false hasContentIssue false
  • English
  • Français

The extraction, characterization and in vitro translation of RNA from adult Schistosoma mansoni

Published online by Cambridge University Press:  06 April 2009

M. P. R. Tenniswood  and
A. J. G. Simpson
M. P. R. Tenniswood
Affiliation:
Laboratory of Developmental Biochemistry, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7
A. J. G. Simpson
Affiliation:
Laboratory of Developmental Biochemistry, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7
Get access Rights & Permissions [Opens in a new window]

Summary

We have extracted RNA from Schistosoma mansoni using the lithium chloride–urea method which gives good yields of undegraded RNA. The results of agarose gel electrophoresis of RNA extracted by this procedure suggest that S. mansoni has an in vivo nick in the large rRNA sub-unit. Translation of the RNA in a rabbit reticulocyte lysate gave significant incorporation of [35S]methionine into synthesized proteins. Immuno-precipitation of these translation products using a hyperimmune monkey serum sedimented between 5 and 8% of the radioactivity, which appeared to be present in approximately 13 proteins of molecular weights 18, 20, 21, 22, 23, 35, 40, 54, 60, 70, 74, 78 and 105 K Daltons.

Type
Research Article
Information
Parasitology , Volume 84 , Issue 2 , April 1982 , pp. 253 - 261
Copyright
Copyright © Cambridge University Press 1982

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Applebaum, S. W., James, T. C., Wreschner, D. H. & Tata, J. R. (1981). The preparation and characterisation of locust vitellogenin messenger RNA and the synthesis of its complementary DNA. The Biochemical Journal 193, 209–16.CrossRefGoogle ScholarPubMed
Auffray, C. & Rougeon, F. (1980). Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. European Journal of Biochemistry 107, 303–14.CrossRefGoogle ScholarPubMed
Bonner, W. M. & Laskey, R. A. (1974). A film detection for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46, 83–8.CrossRefGoogle ScholarPubMed
Capron, A., Auriault, C., Mazingue, C., Capron, M. & Torpier, G. (1980). Schistosome mechanisms of invasion. In The Host-Invader Interplay (ed. Van den Bossche, H.), pp. 217–225Amsterdam: Elsevier/North Holland Press.Google Scholar
Cesari, I. M., Simpson, A. J. G. & Evans, W. H. (1981). Properties of a series of tegumental membrane-bound phosphohydrolytic activities of Schistosoma mansoni. The Biochemical Journal (in the Press).CrossRefGoogle Scholar
Clegg, J. A. & Smithers, S. R. (1972). The effects of immune rhesus monkey serum on schistosomula of Schistosoma mansoni during cultivation in vitro. International Journal for Parasitology 2, 79–98.CrossRefGoogle ScholarPubMed
Cordingley, J. S. & Turner, M. J. (1980). Isolation and characterisation of polysomes from Trypanosoma brucei. Parasitology 81, 537–51.CrossRefGoogle ScholarPubMed
Cox, R. A. (1977). Structure and function of prokaryotic and eukaryotic ribosomes. Progress in Biophysics and Molecular Biology 32, 193–231.CrossRefGoogle ScholarPubMed
Darnborough, C., Legon, S., Hunt, T. & Jackson, R. J. (1973). Initiation of protein synthesis: Evidence for messenger RNA-independent binding of methionyl-transfer RNA to the 40S ribosomal subunit. Journal of Molecular Biology 21, 29–41.Google Scholar
Dunham, S. M., Tenniswood, M. P. R. & Tata, J. R. (1981). Is selection of poly(A) + RNA necessary for establishing a cDNA clone bank? Abstract at 14th FEBS meeting,Edinburgh, Scotland.CrossRefGoogle Scholar
Gilbert, W. & Villa-Komaroff, L. (1980). Useful proteins from recombinant bacteria. Scientific American 242, 68–84.CrossRefGoogle ScholarPubMed
Hoeijmakers, J. H. J., Borst, P., Van den Berg, J., Weissmann, C. & Cross, G. A. M. (1980). The isolation of plasmids containing DNA complementary to messenger RNA for variant surface glycoproteins of Trypanosoma brucei. Gene 8, 391–417.CrossRefGoogle ScholarPubMed
Ishikawa, H. (1976). The fragments from the 28S ribosomal RNA of Galleria mellonella with unesterified uridine at the 3′-termini. Biochemica et biophysica acta 425, 185–95.CrossRefGoogle ScholarPubMed
James, T. C. & Tata, J. R. (1980). Messenger RNA during early embryogenesis in Artemia salina. Altered translatability and sequency complexity. Differentiation 16, 9–21.CrossRefGoogle Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, London 227, 680–5.CrossRefGoogle ScholarPubMed
Lane, C. D., Colman, A., Mohun, T., Morser, J., Champion, J., Kourides, I., Craig, R., Higgins, S., James, T. C., Applebaum, S. W., Ohlsson, R. I., Paucha, E., Houghton, M., Matthews, J. & Miflin, B. J. (1980). The Xenopus oocyte as a surrogate secretory system. The specificity of protein export. European Journal of Biochemistry 111, 225–35.CrossRefGoogle ScholarPubMed
Lukacs, J., Tanaka, R. D., Kim, R. A. & MacInnis, A. J. (1980). Development of cell-free protein synthesising system from Schistosoma mansoni and a comparison of the effects of trycanthone and praziquantel on this system. Journal of Parasitology 66, 424–7.CrossRefGoogle Scholar
Marcu, K. B., Valbuena, O. & Perry, R. P. (1978). Isolation, purification and properties of mouse heavy-chain immunoglobulin mRNAs. Biochemistry 17, 1723–33.CrossRefGoogle ScholarPubMed
McMaster, G. K. & Carmichael, G. G. (1977). Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proceedings of the National Academy of Sciences USA 74, 4835–8.CrossRefGoogle ScholarPubMed
Minty, A. J. & Gros, F. (1980). Coding potential of non-adenylated messenger RNA in mouse friend cells. Journal of Molecular Biology 61–84.CrossRefGoogle Scholar
O'Malley, B. W., Roop, D. R., Lai, E. C., Nordstrom, J. L., Catterall, J. F., Swaneck, G. E., Colbert, D. A., Tsai, M.-J., Dugaiczyk, A. & Woo, S. L. C. (1979). The ovalbumin gene, organisation, structure, transcription and regulation. Recent Progress in Hormone Research 35, 1–42.Google ScholarPubMed
Pelham, H. R. B. & Jackson, R. J. (1976). An efficient mRNA-dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67, 247–56.CrossRefGoogle ScholarPubMed
Rhoads, R. E. (1975). Ovalbumin messenger ribonucleic acid. Purification and fractionation on the basis of polyadenylate content by thermal elution from oligodeoxythymidylate-cellulose. Journal of Biological Chemistry 250, 8088–97.CrossRefGoogle ScholarPubMed
Roskam, W. G. & Rougeon, F. (1979). Molecular cloning and nucleotide sequence of human growth hormone structural gene. Nucleic Acids Research 7, 305–20.CrossRefGoogle ScholarPubMed
Rotmans, J. P. & Vinkestijn, M. (1980). Analysis of the humoral immune response to excretion and secretion antigens of Schistosoma mansoni in infected humans and mice and in immunised rabbits. In The Host-Invader Interplay (ed. Van den Bossche, H.), pp. 151–154. Amsterdam: Elsevier/North Holland Press.Google Scholar
Simpson, A. J. G., Schryer, M. D., Cesari, I. M., Evans, W. H. & Smithers, S. R. (1981). Isolation and partial characterization of the tegumental outer membrane of adult Schistosoma mansoni. Parasitology 83, 163–77.CrossRefGoogle ScholarPubMed
Smithers, S. R. & Terry, R. J. (1965). Infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms. Parasitology 55, 695–700.CrossRefGoogle ScholarPubMed
Snary, D., Smith, M. A. & Clegg, J. A. (1980). Surface proteins of Schistosoma mansoni and their expression during morphogenesis. European Journal of Immunology 10, 573–5.CrossRefGoogle ScholarPubMed
Tata, J. R. & Baker, B. (1975). Sub-nuclear fractionation. III. Sub-nuclear distribution of poly(A)-rich RNA. Experimental Cell Research 93, 191–201.CrossRefGoogle ScholarPubMed
Taylor, D. W., Hayunga, E. G. & Vannier, W. E. (1981). Surface antigens of Schistosoma mansoni. Molecular and Biochemical Parasitology 3, 157–68.CrossRefGoogle ScholarPubMed