![]() From Gottfried Schatz comment on (unpublished?) data at the end of this intro to mt in Ann Rev Biochem 2007: "the two proteins encoded by all mitochondrial DNAs, subunit I of cytochrome oxidase and cytochrome b, are precisely those that are exceptionally hydrophobic. Is there anything to be learned if - result seen w/o tag? Hydrophobicity predictor tool here. may want to order this sequence only since import to mt of bulky HA or charge myc may be problematic.NotI is missing from deoxycycline regulatory module and is present as unique site in polylinker of all pRS series. other sites not in mtRNT1 that are in pRS416 polylinker after RI, Bam pair: SpeI, XbaI, NotI, EagI, AleI, SacII, BstXI, SacI.sequence file shows this variation does not have either an EcoRI or a BamHI site.removed AAG AAT AAG AAA AGA AAA TTC TCA GAT ACA AGC TGA/Lys Asn Lys Lys Asn Lys Phe Ser Asp Thr Ser, last 11 aa before stop codon, keeping stop codon.described as sufficient for nuclear localization of RNT1.removed nuclear localization signal from 3'end fo RNT1 gene ![]() sequence ATG CAA CGC TCC ATT TTT GCG AGG TTC/ Met Gln Arg Ser Ile Phe Ala Arg Phe.does not change restriction analysis with favorite enzymes.described in PMID: 3023841 as sufficient for mt localization of b-gal.added mitochondrial targeting sequence from HEM1 to 5' end of RNT1 Changed GA A(bp721-723)TTC(bp724-726) at Glu240Phe241 to GA G TTC which removed RI but keeps GluPhe. removed EcoRI site from coding sequence of RNT1. RNT1/YMR239C on chromosome XIII from coordinates 749676 to 748261 on Crick (bottom) strand. downloaded genomic sequence of RNT1 from SGD. Steps to target RNT1 to mitochondrial matrix Not ordered since cloned tTA/CYCI into pRS416 on RI/Bam frag. sequence file for tTA+CYC1tetO2+mtRnt1+HA. sequence file for tTA+CYC1tetO2 from pCM224, RI to Bam unique BamHI at 5' end, unique NotI at 3'end.sequence file for synthesis order = mtRNT1 with: MtGFP= JSC350X and parent strain DL2: 070307 strain from Tom Fox, 070607 photos from Matt Gethers. ![]() deoxycycline-regulation of RNase expression
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