User talk:Nkuldell/mtDNA pt3: Difference between revisions
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==deoxycycline-regulation of RNase expression== | ==deoxycycline-regulation of RNase expression== | ||
[[Image:Macintosh HD-Users-nkuldell-Desktop-pCM224.PNG.PNG|thumb|plasmid map for pCM224]] | [[Image:Macintosh HD-Users-nkuldell-Desktop-pCM224.PNG.PNG|thumb|plasmid map for pCM224]] | ||
*regulation of expression from deoxycline-induced promoter, described in [http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=9778798&ordinalpos=4&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Herrero Yeast 1998 14(12):1127-38] | |||
*[[Media:Macintosh HD-Users-nkuldell-Desktop-mtRNA silencing-pCM224seq.rtf| sequence file]] for pCM224 which encodes tTA and CYC1-repressible promoter due to 2 copies of tetO | |||
* <b>can excise relevant regulatory module (tTA driven by ADH1 and CYC1 with 2XtetO) on RI, BamHI fragment</b> | |||
==pRS416== | ==pRS416== | ||
[[Image:Macintosh_HD-Users-nkuldell-Desktop-pRS416singlecutters.PNG.PNG|thumb|left| plasmid map for pRS416 with single cutters indicated]] | [[Image:Macintosh_HD-Users-nkuldell-Desktop-pRS416singlecutters.PNG.PNG|thumb|left| plasmid map for pRS416 with single cutters indicated]] | ||
*[[Media:PRS416_fromNEB.rtf| sequence file]] for pRS416 | |||
* <b>can clone tTA/CYC1tetO2/mtRNT1 into RI, BamHI bkb</b> | |||
==mtRnt1== | ==mtRnt1== | ||
Steps to target RNT1 to mitochondrial matrix | |||
# downloaded genomic sequence of RNT1 [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=rnt1 from SGD]. RNT1/YMR239C on chromosome XIII from coordinates 749676 to 748261 on Crick (bottom) strand. | |||
*[[Media:Macintosh HD-Users-nkuldell-Desktop-RNT1 genomic fromSGD.rtf| sequence file]] for RNT1 genomic DNA | |||
Revision as of 11:49, 26 June 2007
Experimental Details
deoxycycline-regulation of RNase expression
- regulation of expression from deoxycline-induced promoter, described in Herrero Yeast 1998 14(12):1127-38
- sequence file for pCM224 which encodes tTA and CYC1-repressible promoter due to 2 copies of tetO
- can excise relevant regulatory module (tTA driven by ADH1 and CYC1 with 2XtetO) on RI, BamHI fragment
pRS416
- sequence file for pRS416
- can clone tTA/CYC1tetO2/mtRNT1 into RI, BamHI bkb
mtRnt1
Steps to target RNT1 to mitochondrial matrix
- downloaded genomic sequence of RNT1 from SGD. RNT1/YMR239C on chromosome XIII from coordinates 749676 to 748261 on Crick (bottom) strand.
- sequence file for RNT1 genomic DNA
