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Evidence for Virus Population Heterogeneity in the Sheep and Goat Pox
Vaccine Strain 0240
Sheep and goat
pox vaccine strain 0240 was characterized. We designed a pair of primers
flanking 3 ORFs including thymidine kinase (tk) gene of Kenya sheep-1.
Polymerase chain reaction (PCR) was performed and PCR products of
approximately 908 base pair (bp) in size were obtained for all cloned
plaques and the vaccine strain. Lamb testicle (LT) DNA (negative
control) as expected did not produce any PCR products. PCR products were
digested with Ssp1 and Dra1 restriction enzymes and analyzed using
agarose gel and polyacrylamide gel electrophoresis (PAGE), respectively.
Generation of the expected banding pattern following digestion confirmed
virus identity. The analysis revealed the presence of restriction
fragment length polymorphisms (RFLPs) between some plaques at
temperatures of 33°C and 37°C and the vaccine strain. An in vitro
recombination experiment was performed for the 0240 vaccine strain,
which revealed its viral sequence heterogeneity. The pattern of heat
inactivation indicated the presence of at least two virus populations
differing in heat sensitivity. In conclusion, these experiments
indicated the presence of two different virus populations. |
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