Part:BBa_K1405004

McfR

Background

McfR is a TCA intermediates responsive chemoreceptor that originally found in ''Pseudomonas putida''. BBa_K1405004 contains McfR coding sequence, an insulator, RBS sequence, and the constitutive promoter BBa_J23100, which is similar to BBa_K515102 that can help us compare our work with iGEM11_Imperial_College _London had done. And we showed the results below. A 15 bp insulator sequence upstream of the RBS ensures tunability of expression through easy switching of promoters. In addition it allows the translation initiation rate (TIR) of the RBS to remain the same, when the promoter is replaced.This device is compatible with motile strains of E. coli. It has been transformed and tested in E. coli BL21 in the vector backbone pSB1C3. This device is used as an additional chemoreceptor for endogenous chemotaxis system in E. coli and responds to L(-) malic acid (HO2CCH2CHOHCO2H) and succinate (HOOC-(CH2)2-COOH).

Result

Figure 1 Expression and purification of ModA. Expression of ModA and its purification by His-chelating chromatography. Lane 1, molecular weight standards (kDa); lane 2, total bacterial proteins before IPTG induction; lane 3, total bacterial proteins after 0.5 mM IPTG induction; lane 4, total bacterial lysate after 0.5 mM IPTG induction; lane 5,the sediment after 16,000 rpm 30 min; lane 6 the flow through of column loading; lane 7,the flow of buffer B .lane 8 The purification of ModA by size-elution chromatography.

Figure 1 Expression and purification of ModA. Expression of ModA and its purification by His-chelating chromatography. Lane 1, molecular weight standards (kDa); lane 2, total bacterial proteins before IPTG induction; lane 3, total bacterial proteins after 0.5 mM IPTG induction; lane 4, total bacterial lysate after 0.5 mM IPTG induction; lane 5,the sediment after 16,000 rpm 30 min; lane 6 the flow through of column loading; lane 7,the flow of buffer B .lane 8 The purification of ModA by size-elution chromatography.

Figure 2 Native-PAGE to provide the molybdate lane 1, the modA(acetic buffer PH 5.0); lane 2, the modA incubate with 20mM molybdate.

Sequence and Features BBa_K1405004 SequenceAndFeatures