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Construction of a nonpolar disruption in pstB. An internal fragment of the pstB gene (a and b) was cloned into pBGST18 (a′ and b′) in the same orientation as the lacZ promoter of the plasmid (Plac). Integration of the plasmid by a single crossover generates two truncated copies of pstB. pstB′ is missing C-terminal coding sequences, and ′pstB is missing N-terminal coding sequences. The Plac promoter of the plasmid ensures transcription of the downstream genes.
