Soluble expression, purification and characterization of the full length IS2 Transposase

Table 1 Distribution, from 23 recovered mutants, of 25 randomly induced mutations in the four domains of the IS2 OrfAB TPase

^aDomains of the IS2 OrfAB TPase, deduced from these studies are as follows: H + HTH: the Binding domain; LZ-L: Leucine zipper-like oligomerization motif; MI: the middle interval; CAS: putative catalytic active site which extends from residue 236-398. The locations of the three catalytic carboxylates are shown. The sequence associated with D240 includes the β strands 1-3, that with D306, β strands 4-5 and α helices 1-3 and that with E342 α helices 4-6. Intervals represented by the numbers 1-14, 61-69, 207-235, and 399-409 are most likely disordered sequences (see Figure 9a). ^bThe wild type fusion protein (OrfAB-GFP) was overexpressed from the plasmid pLLIS2orfAB::GFP (pLL2522). Mutant proteins were overexpressed from the GMF strains carrying similar plasmids (pLL2524-XXX i.e., from 001-110) carrying a mutagenized orfAB gene. Isolated GMF strains were numbered from 1-110.

ISSN: 1759-8753