[Identification and application of yeast histone acetyltransferases Elp3 polyclonal antibody].
Yeast Elongation protein 3 (yElp3), the catalytic subunit of the multi-subunit histone acetyltransferase elongator complex, is involved in histone acetylation and transcription, exocytosis and tRNA modification. To study the complex function of yElp3 in yeast, we amplified the yElp3 gene fragment encoding 73aa in the N-terminal from plasmid pYES2-yElp3, and then cloned it into pMXB10 to construct the recombinant plasmid pMXB10-yElp3-219. We expressed the fusion protein in E. coli BL21 (DE3), then purified it by chin affinity column, and finally obtained the soluble purified protein (8.0 kD), which was used to immune the rabbits for acquiring antiserum. ELISA and Western blotting indicated that the polyclonal antibody was of high titration and specificity. Chromatin immunoprecipitation (ChIP) assay with this antibody suggested that yhElp3 exerted the transcriptional regulatory function directly through its presence on the SSA3 gene; this might be the reason that it can rescue the delay activation of SSA3 in elp3delta cells.
Li
Fen
F
College of Life Sciences, Henan Normal University, Xinxiang 453007, China. lifen2001@yahoo.com
Tian
Shujuan
S
Zhang
Shuai
S
Kong
Yan
Y
Wang
Yanfang
Y
D000595:Amino Acid Sequence; D000906:Antibodies / Q000032:analysis / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D051548:Histone Acetyltransferases / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C526363:Elp3 protein, S cerevisiae; D051548:Histone Acetyltransferases