Cm ori
 & uncB (a)
 E (c)

 ~F (b+11, cys20-mer, gly43-wys

 H (8cysl40-mer)


 p TA M24
 10.9 kb



 G (y



 C (s)

Figure 4-3. Expression plasmid of cysteine mutants. F1Fo ATP synthase complexes with
 site-specific cysteine mutations were designed to be expressed from a single
 plasmid encoding the entire unc operon. Plasmid pTAM24 is one of twelve of
 the constructed plasmids. The name of the unc gene is listed followed by the
 subunit it expresses in parenthesis. The black stars represents codons that
 expressed a native cysteine that had been substituted with a serine. The red
 star represents a glycine to cysteine substitution at codon 43 of the uncF(b)
 gene. The green dot represents an 11 amino acid insertion. Plasmid pTAM24
 expresses the entire unZc operon with the following mutations: b+11, cys200ser,
 gly434cys, 6cysl400ser, and acys900ser. The plasmids confer chloramphenicol
 resistance and the pACYAl84 origin of replication. Similar constructions
 include those listed in Table 4-2.

however, it was necessary to determine whether the mutations generated in the present

study would have affect activity. The effects of the cysteine mutations were studied by

the ability of the plasmids to complement either the E. coli strain KM2 (Ab) (Table 4-1)

or 1100ABC (BApyisabc) (Table 4-3). Growth on succinate minimal medium was used