His workStrains/plasmids Bacteria R. capsulatus MT-SRP1.r1 MD11 MD14 E.
His workStrains/plasmids Bacteria R. capsulatus MT-SRP1.r1 MD11 MD14 E. coli HB101 XL1 Blue BL21(DE3) Plasmids pAV1 pAV10 Relevant propertiesa Refs.(ccmI::kan) G488A up-promoter of ccmFH; Res Ps (ccmG::kan); Res Ps ccmH::spe; Res Ps F (gpt-proA)62 araC14 leuB6(Am) glnV44(AS) thi-1 galK2(Oc) lacY1 (mcrC-mrr) rpsL20(Strr) xylA5 mtl-1 endA1 gyrA96(NalR) thi-1 recA1 relA1 lac glnV44 F [::Tn10 proAB lacIq (lacZ)M15] hsdR17(rK mK ) fhuA2 [lon] ompT gal ( DE3) [dcm] hsdS DE3 sBamHI o EcoRI-B int:: (lacI::PlacUV5::T7 gene1) i21 nin50 51 50 Stratagene Stratagene New England Biolabs 29 This work This function This perform This operate Stratagene This perform This function This operate This function This function This operate This perform 51 This operate 31 29 30 This perform This perform 30 51 QiagencpCS1302 derivative carrying truncated R. capsulatus cycA at the NdeI amHI web-sites using a Strep-tag II at its five -end and without a IFN-gamma Protein Species signal sequence, AmpR (Strep-apocyt c2WT)b pFLAG-1 derivative carrying truncated R. capsulatus ccmH lacking its signal sequence and TM helix (C-terminal 46 amino acids) from pCS1604 at the NdeI amHI internet sites, AmpR (FLAG-CcmHWT) pAV10C42S pAV10 derivative with C42S mutation in R. capsulatus ccmH, AmpR (FLAG-CcmHCys-45) AV10C45S pAV10 derivative with C45S mutation in R. capsulatus ccmH, AmpR (FLAG-CcmHCys-42) pAV10C42SC45S pAV10 derivative with C42S and C45S mutations in R. capsulatus ccmH, AmpR (FLAG-CcmH) pBSK AmpR pCS1552 pQE60-helX derivative with C75S mutation in R. capsulatus ccmG, AmpR pCS1553 pQE60-helX derivative with C78S mutation in R. capsulatus ccmG, AmpR pCS1554 pQE60-helX derivative with C75S and C78S mutations in R. capsulatus ccmG, AmpR pCS1555 A truncated R. capsulatus ccmG from pQE60-helX cloned in to the EcoRI indIII web sites of pBSK, AmpR (His6 cmGWT) pCS1556 A truncated R. capsulatus ccmG with C75S mutation taken from pCS1552 and cloned into the EcoRI indIII websites of pBSK, AmpR (His6-CcmGCys-78) pCS1557 A truncated R. capsulatus ccmG with C78S mutation taken from pCS1553 and cloned into the EcoRI indIII web-sites of pBSK, AmpR (His6-CcmGCys-75) pCS1558 A truncated R. capsulatus ccmG with C75S and C78S mutations, taken from pCS1554 and cloned into the EcoRI indIII web sites of pBSK, AmpR (His6-CcmG) pCS1566 pCHB500 derivative producing R. capsulatus ccmG with its signal peptide and His6-tagged at its 3 -end, cloned in to the XbaI/ KpnI web-sites, TetR (His6-CcmGWT) pCS1604 pACCyc3 derivative (52), creating a truncated R. capsulatus ccmH from Endosialin/CD248 Protein Storage & Stability pRGK291 (13), lacking both its signal sequence and TM helix (final 46 amino acids at its 3 -end), cloned into NdeI-BamHI web sites. ClmR pFLAG-1 pMADO5 derivative carrying five -FLAG-tag ccmI (as opposed to His10 tag), AmpR pMADO5 pCS1303 derivative, containing R. capsulatus ccmI carrying 5 -His10 tag, AmpR (His10-CcmIWT) pMAM1 pCS1302 derivative, containing truncated R. capsulatus petC (derivative of petC with no signal sequence and lacking its last C-terminal 39 amino acids, and its non-heme ligating Cys-144 and Cys-167 residues) cloned into the NdeI amHI sites with a Strep-tag II fused at its five -end, AmpR (Strep-apocyt c1WT) pMAM1C34S pMAM1 derivative with C34S mutation of R. capsulatus petC, AmpR (Strep-apocyt c1Cys-37) pMAM1C37S pMAM1 derivative with C37S mutation of R. capsulatus petC, AmpR (Strep-apocyt c1Cys-34) pMAM1C34SC37S pMAM1 derivative with C34S and C37S mutations of R. capsulatus petC, AmpR (Strep-apocyt c1) pNJ2 pRK415 derivative making R. capsulatus five -end FLAG-tagged ccmI with its personal promoter, TetR (FLAG-CcmIWT) pQE60.
