Fig. 3. Functional analysis of ssrA^Ng alleles with transposon insertions in and surrounding ssrA^Ng. A library of 13 m-Tncm insertions was created in plasmid pGCtm. The positions (shown by downward pointing arrows) in the GCtm fragment of these insertions were located by DNA sequencing. Insertions were located in three regions: A, upstream of ssrA; B, within ssrA; and C, downstream of ssrA. The effect on tmRNA activity of each insertion was determined by assessing growth of λimmP22hy25 in derivatives of the ssrA^Ec::cat E.coli strain K8619 carrying variants of pGCtm with the respective 13 insertions (row 1); (+) support of phage growth and (–) no support of phage growth. Formation of stable recombinants with mini-transposon inserted ssrA^Ng alleles in N.gonorrhoeae either haploid (row 2) or diploid (row 3) for ssrA^Ng was determined by selecting Cm^r colonies; (+) Cm^r colonies were isolated and (–) Cm^r colonies were not isolated.