MOLECULAR CHARACTERIZATION OF CITRUS YELLOW MOSAIC VIRUSB.V. B. REDDY, Y.S. AHLAWAT AND ANUPAM VARMA
Nucleic acid extracted from purified virus was unaffected by RNase A and Si nuclease treatment but completely degraded by DNase A treatment indicating that the viral genome is ds DNA. In nondanaturing agarose gels the viral DNA migrated as two bands at 21Kb and 6.6 Kb . The faster migrating band of 6.6 Kb may be linear from of viral DNA while slow migarating band of 21Kb may be circular form of viral DNA by anology with other badnaviruses indicating that viral genome is circular in nature. Digestion of viral DNA with Si nuclease gave two extra bands which are absent in untreated DNA indicating that the viral genome has two single stranded discontinuities may be one on each strand. The genome size of virus was estimated as 7.2 Kb (approx.) by Si nuclease treatment . The viral genome was cloned in pUC18 at Hind III site and identified two virus specific clones of 2.0 Kb and 4.0Kb in length by dot blot hybridization and their restriction maps were prepared. Anlaysis of capsid protein of virus by SDS-PAGE and Western blot gave one major capsid protein of 32 Kd and two minor bands of 22 and 19 Kd which are rather degraded products of 32 Kd.
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