Abstract
Publisher Summary This chapter describes the construction of NP–BPV recombinant DNA and isolation of such recombinant DNA transformants that produce influenza virus NP. To construct NP–BPV recombinant DNA, the Bam HI cleaved full-length NP DNA was inserted into the Bgl II site located between the MT promoter and the SV40 polyA addition site of cloned vector DNA containing the pML2 moiety (a derivative of pBR322). The recombinant DNA containing the NP DNA inserted in the sense transcription orientation was selected and partially digested to open either of the two remaining Bam HI sites: one located within the NP gene and the other located downstream of the SV40 polyA addition sequence. The NP–BPV recombinants that contained BPV DNA inserted at the Bam HI site located downstream of the SV40 polyA site were selected. The chapter illustrates the final construct of NP–BPV DNA in which the transcription orientation of the BPV insert is opposite to the transcription direction of the NP gene.
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