Fig. 2.

Properties of SgrS variants possessing short polyU tails. (A) Nucleotide sequences of the rho-independent terminators of SgrS variants. The inverted repeat sequences are underlined. The sequences are based on the assumption that the transcription of the corresponding genes terminates at the last U within the polyU tail. (B) Effects of shortening of polyU tail on the expression and function of SgrS. IT1568 and TM589 cells harboring indicated plasmids were grown in LB medium. At A₆₀₀ = 0.6, total RNAs were prepared and each RNA sample was subjected to Northern blot analysis using the SgrS probe 1 and the ptsG probe. tmRNA was used as a loading control. The following amounts of RNAs were loaded: SgrS variants, 0.25 μg; the ptsG mRNA, 7 μg; tmRNA, 0.25 μg. Total proteins were also prepared and samples equivalent to 0.025 A₆₀₀ units were subjected to Western blot analysis using anti-IIB antibodies.