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Characterization of germiston virus rna molecular weight and oligo nucleotide fingerprints of the 3 rna species

Characterization of germiston virus rna molecular weight and oligo nucleotide fingerprints of the 3 rna species

Annales de Virologie (Paris) 133(4): 461-472

The 3 genomic RNA segments of the Germiston virus (Bunyaviridae) were analyzed by electrophoresis in denaturing gels. A variety of gel types and denaturing agents were tested. Best separation of the 3 RNA species was achieved using 1% agarose gels in the presence of methylmercuric hydroxide. Under these conditions, the L, M and S RNA segments migrated with apparent MW of 2.8 .times. 106, 1.6 .times. 106 and 0.34 .times. 106 daltons, respectively. The RNA species were isolated from the gel using low-melting temperature agarose after which they were digested with RNase-T1, and the resulting oligonucleotides were labeled in vitro using T4 polynucleotide-kinase and .gamma.32P-ATP. The RNase-T1-resistant oligonucleotide fingerprints obtained after 2-dimensional electrophoresis show that each of the 3 viral RNA segments is unique.

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