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Nucleic Acids Research, 1991, Vol. 19, No. 7 1613-1618
© 1991

MOLECULAR BIOLOGY

Hybridization arrest of the cell-free translation of the malarial dihydrofolate reductase/thymidylate synthase mRNA by anti-sense oligodeoxyribonucleotides

Christian Sartorius and Richard M. Franklin*

Biocenter, Department Structural Biology Klingelbergstrasse 70, CH-4056 Basel, Switzerland

* To whom correspondence should be addressed

Received November 20, 1990. Revised March 7, 1991. Accepted March 7, 1991.

In order to inhibit the in vitro translation of Plasmodium falciparum mRNA coding for the bifunctional enzyme dihydrofolate reductase-thymidylate synthase (DHFR-TS), oligodeoxynucleotides (ODNs) were directed against the translation initiation site or a site in the TS-coding region. In both cases considerable hybridization arrest, i.e.>50% inhibition, was only achieved if the lengths of the ODNs to the two regions were 30 and 39 nucleotides, respectively, or longer. The ODN with the highest efficiency was a 49-mer directed against the TS-coding region (OTS49); 45 µM was sufficient to inhibit the expression of DHFR-TS by almost 90%. In this case the synthesis of DHFR-TS was interrupted at the binding site of OTS49 by a RNase H-independent mechanism. The resulting polypeptide was smaller (55 kDa) than one subunit of the native protein (71 kDa) and lacked TS activity.


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