发明名称 |
Mutants Of O6-Alkylguanine-Dna Alkyltransferase |
摘要 |
The invention relates to AGT mutants showing, when compared to the wild type human AGT, two or more advantageous properties selected from (a) reduced DNA interaction; (b) localisation of the expressed protein in eukaryotic cells that is no longer restricted to the nucleus; (c) improved expression yield as soluble protein and improved stability in various hosts; (d) improved stability under oxidising conditions; (e) improved stability within cells after reaction with a substrate; (f) improved stability outside cells before and after reaction with a substrate; (g) improved in vitro solubility; (h) improved reactivity against 0<SUP>6</SUP>-alkylguanine substrates; (1) reduced reactivity against DNA-based substrates; and (j) reduced reactivity against N<SUP>9</SUP>-substituted 0<SUP>6</SUP>-alkylguanine substrates. Such AGT mutants with the mentioned improved properties are mutants wherein between 1 and 25 amino acids of the wild type human AGT are substituted by other amino acids, and optionally I to 5 amino acids out of the continuous chain at one, two or three positions are deleted or added and/or 1 to 4 amino acids at the N-terminus or 1 to 40 amino acids at the C-terminus are deleted. The invention further relates to a method for detecting and/or manipulating a protein of interest wherein the protein of interest is incorporated into a fusion protein with the AGT mutants of the invention. Another object of the invention are AGT fusion proteins comprising such AGT mutants and the protein of interest.
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申请公布号 |
US2007207532(A1) |
申请公布日期 |
2007.09.06 |
申请号 |
US20050591159 |
申请日期 |
2005.03.01 |
申请人 |
BARNIKOV JAN;CHIDLEY CHRISTOPHER;GRONEMEYER THOMAS;HEINIS CHRISTIAN;JACCARD HUGHES;JOHNSSON KAI;JUILLERAT ALEXANDRE;KEPPLER ANTJE |
发明人 |
BARNIKOV JAN;CHIDLEY CHRISTOPHER;GRONEMEYER THOMAS;HEINIS CHRISTIAN;JACCARD HUGHES;JOHNSSON KAI;JUILLERAT ALEXANDRE;KEPPLER ANTJE |
分类号 |
C12N9/00;C12N9/10 |
主分类号 |
C12N9/00 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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