User talk:Nkuldell/SAGA swap pt 2
SGF73 emailed info
From: Mark Hickman Subject: Re: memory lapses... Date: Sun, 11 Jun 2006 21:32:42 -0400
SGF73 is required for the induction of the PAU genes under hypoxic conditions. As far as I know, this is the first transcriptional defect seen for sgf73 mutants. Otherwise, nothing is known about its function. Dom worked on human Sgf73 for his PhD thesis; he found it was subject to glutamine expansion. One of my plans for the future was to check whether sgf73 deletion has a phenotype, especially under hypoxic conditions (since there might be a consequence to the PAU genes not being induced). Another idea was to do a synthetic lethal screen with sgf73 under aerobic and hypoxic conditions.
We have the cerevisiae sgf73 deletion strain (I have worked with FY2475). Not sure if there is a cDNA, but Fred or Lisa might know.
Let me know if you have any other questions. You could also ask Dom about pombe sgf73+. He might be knocking that out and/or have a cDNA.
Thanks for the anniversary wishes! It's July 31. I guess we are still newlyweds until the first year?
Good luck with your son's surgery. I have a memory lapse too: your son's name.
See you soon Mark
From: Mark Hickman Subject: Re: memory lapses... Date: Fri, 23 Jun 2006 11:25:35 -0400
No problem! Just got back from Venice. How did it go with Spencer?
It's MAT alpha ura3-52 lys2-173R2 leu2D1 arg4-12 sgf73D0::kanMX his4-917d. I am assuming it's a clean ORF deletion but not sure. It should be in Jenny's notebook, or maybe Lisa knows? Won't you need to put in URA3, so you can do the swap with 5FOA?
Do you know about the Winston Lab yeast strains on the web? http://genepath.med.harvard.edu/strainfinder/yeast.php
Lisa is still in the lab. Not much longer though....
From: Mark Hickman Subject: Re: memory lapses... Date: Fri, 23 Jun 2006 11:49:41 -0400
Natalie, I just happened to come across a file (totally randomly!) with Jenny's knockout primers: Sgf73 JW109 left primer 5' TGAACACACAAGAGAAGCGCAAAAGAGTAAAGAGCTAAA ctgtgcggtatttcacaccg JW110 right primer 5' CTCACTTCGTGAACATGCTGGATAACGTGCATGATTCAA agattgtactgagagtgcac
URA3 from pRS416 according to NEB site
URA3 from SGD is 804 bp vs this gene that weighs in at 1213 bases. SGD BLAST gives perfect match to SGD URA3 sequence from 1-804
1 A TGTCGAAAGC TACATATAAG GAACGTGCTG CTACTCATCC
451 TAGTCCTGTT GCTGCCAAGC TATTTAATAT CATGCACGAA AAGCAAACAA
501 ACTTGTGTGC TTCATTGGAT GTTCGTACCA CCAAGGAATT ACTGGAGTTA
551 GTTGAAGCAT TAGGTCCCAA AATTTGTTTA CTAAAAACAC ATGTGGATAT
601 CTTGACTGAT TTTTCCATGG AGGGCACAGT TAAGCCGCTA AAGGCATTAT
651 CCGCCAAGTA CAATTTTTTA CTCTTCGAAG ACAGAAAATT TGCTGACATT
701 GGTAATACAG TCAAATTGCA GTACTCTGCG GGTGTATACA GAATAGCAGA
751 ATGGGCAGAC ATTACGAATG CACACGGTGT GGTGGGCCCA GGTATTGTTA
801 GCGGTTTGAA GCAGGCGGCA GAAGAAGTAA CAAAGGAACC TAGAGGCCTT
851 TTGATGTTAG CAGAATTGTC ATGCAAGGGC TCCCTATCTA CTGGAGAATA
901 TACTAAGGGT ACTGTTGACA TTGCGAAGAG CGACAAAGAT TTTGTTATCG
951 GCTTTATTGC TCAAAGAGAC ATGGGTGGAA GAGATGAAGG TTACGATTGG
1001 TTGATTATGA CACCCGGTGT GGGTTTAGAT GACAAGGGAG ACGCATTGGG
1051 TCAACAGTAT AGAACCGTGG ATGATGTGGT CTCTACAGGA TCTGACATTA
1101 TTATTGTTGG AAGAGGACTA TTTGCAAAGG GAAGGGATGC TAAGGTAGAG
1151 GGTGAACGTT ACAGAAAAGC AGGCTGGGAA GCATATTTGA GAAGATGCGG
1201 CCAGCAAAAC TAA