Hexa-His tagged Ivyc,Ivyp, and E. coli MliC proteins were obtained using similar procedures.
Genes coding for Ivyc and E.coli MliC were amplified from E.coli JM105,and that for Ivyp from P.aeruginosa strain PA01. Expression vectors pET26b-Ivyc-his, pET26b-Ivyp-his,and pET26b-MliC-his were constructed and subsequently transformed into Shuffle T7 Express cells(New England Biolabs, USA) and produced as described previously.