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Example sentences for: midasin
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We used regions of amino acid sequence conserved between the midasin genes in yeast and Schizosaccharomyces pombe to make a rough map of the gene onto the August 2001 freeze of the assembled human genome with the public domain Blat server at http://genome.ucsd.edu/index.html [ 54 55 ] . The resultant partial nucleotide sequence identified about four-fifths of the exons and was used to design the requisite PCR primers to verify all exon-intron boundaries in the gene by physical sequencing of appropriate PCR-amplified cDNA fragments.
The 3'-end of the coding region of the midasin gene was fused to a tag encoding 3× hemagglutamin (HA) by using the PCR-mediated cassette procedure of Longtine and colleagues [ 60 ] . The PCR primers used to generate the cassette were 5'-ACTGATTTTG CGTCAATACT TTACAGACCT GGCATCCAGC CGGATCCCCG GGTTAATTAA-3' and 5'-TCGTGTAGTA AACCTCCTCT TCTTGGTTTT CACGATATAC GAATTCGAGC TCGTTTAAAC-3'.
In order to confirm the yeast database entry indicating that the MDN1 (YLR106C) gene is essential for viability in yeast, we performed tetrad analysis of the heterozygous midasin knock-out strain CMEY072(HE) (Mat a/α ura3-52/ura3-52 his3D1/his3D1 leu2-3_112/leu2-3_112 trp1-289/trp1-289; MDN1(4,14727)::kanMX4/MDN1) obtained from EuroScarf.
Adequate migration of the 560 kDa midasin polypeptide on electrophoresis gels requires use of lower percentage gels than standard, especially if blotting is involved.
The AAA domain of midasin is evolutionarily related to that of dynein, but it appears to lack a microtubule-binding site.
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