Strains and plasmids
This protocol is extracted from research article:
A hidden state in the turnover of a functioning membrane protein complex
Sci Adv, Mar 20, 2019; DOI: 10.1126/sciadv.aau6885

All strains used in this study were derived from E. coli K12 strain RP437: JY21 (motA448 ΔfliC ΔcheY), JY27 (ΔfliC ΔcheY), and SM1 (ΔfliC ΔcheY ΔmotB). The plasmid pKAF131 constitutively expresses the sticky filament FliCst. The plasmid pTrc99aMotA expresses wild-type MotA under control of an IPTG-inducible promoter. The plasmid pBAD33MotBΔplug expresses MotBΔ51–70 under control of an arabinose-inducible promoter. The plasmid pFD313 also constitutively expresses the sticky filament FliCst and is compatible with the pBAD33 vector (16). To measure the stator turnover kinetics, the mutant strain JY21 transformed with the plasmids pKAF131 and pTrc99aMotA, JY27 (denoted as the wild-type strain) transformed with the plasmid pKAF131, and SM1 transformed with the plasmids pBAD33MotBΔplug and pFD313 were used.

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