An Examination of the Splicing Event of Intron One in Woodchuck Hepatitis Virus
Pillar, Christopher M.
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This study examines the role of RNA splicing on woodchuck hepatitis virus (WHV) production in vitro. It has previously been determined that two introns exist within the WHV genome (Ogston and Razman, 1992). The acceptor splice consensus sequence of the first intron was mutated so that the normal splicing event of this intron would be theoretically eliminated. From human hepatoma cells transfected with the acceptor site mutants, DNA and RNA were isolated and assayed for virus production. Production of virus comparable to that produced by cells transfected with wild-type WHY DNA was observed with the mutants. To determine whether the splicing event had been successfully eliminated or not, mutant viral DNA was subjected to polymerase chain reaction (peR). Surprisingly, mutating the acceptor splice sequence did not abolish the splicing of the intron. These mutants will be subjected to further investigation, with the hope of identifying how the mutant virus continues to splice even in the presence of a mutated splice sequence.