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Isolation and Control of the Function Quality of Messenger-Rna Bovine Leukocyte Interferon

VILCEK S, FORGAC O, HIPIKOVA V
Veterinarni Medicina 37, 1992, 213-220

For he construction of the cDNA library total cellular RNA was separated from bovine leucocytes induced with NDV [inductor of interferons] for five hours. Guanidinisothiocyanate and ultracentrifugation in the gradient of CsCl were used for the separation of the total RNA [Chirgwin et al., 1979]. Bands 18S and 28S were detected in the samples of RNA by electrophoresis in an MOPS buffer [Fig. 1]. mRNA was separated from the mixture of RNA via affinity chromatography on oligo[dT]-cellulose [Aviv and Leder, 1972). The function quality of mRNA BoIFN-alpha was verified by microinjection into the oocytes of Xenopus laevis using a microinjector of our own construction (Fig. 2). The microinjector was calibrated by injection of a radioactive Cr-51 solution (Fig. 3). It was found out on the basis of CPE inhibition measurements that the injected oocytes synthetized 320-840 U/ml BoIFN-alpha [16-32 U/50-mu-l


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