[Isolation and control of the functional quality of mRNA of bovine leukocyte interferon]. / Izolácia a kontrola funkcnej kvality mRNA bovinného leukocytárneho interferónu.
Vet Med (Praha)
; 37(4): 213-20, 1992 Apr.
Article
em Sk
| MEDLINE
| ID: mdl-1641951
ABSTRACT
For the 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 51Cr solution (Fig. 3). It was found out on the basis of CPE inhibition measurements that the injected oocytes synthesized 320-640 U/ml BoIFN-alpha (16-32 U/50 microliters; Tab. I).
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
RNA Mensageiro
/
Bovinos
/
Interferon-alfa
Limite:
Animals
Idioma:
Sk
Revista:
Vet Med (Praha)
Ano de publicação:
1992
Tipo de documento:
Article