Transposon tagging

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The transposon tagging is a method for identification of known or unknown genes by mutagenesis with transposon .

properties

The random insertion of mobile genetic elements into the genome inactivates genes in unfolded areas of the DNA or - in rarer cases - also activates them. The phenotype changed by the inactivated gene is used to identify the clone mutated in the gene sought . By DNA sequencing with transposon-based primers , the DNA sequence adjacent to the transposon is identified by primer walking . These adjacent DNA sequences can be amplified by a polymerase chain reaction , e.g. B. for a cloning and a subsequent protein purification and protein characterization . If the genome is already completely sequenced, homologies in databases provide information about the known functions of similar DNA sequences ( Blast ) or the resulting proteins ( Pfam ).

Alternative methods are targeting induced local lesion in genomes ( TILLING ) and genome-wide hybridization methods after random mutagenesis.

Individual evidence

  1. ↑ D. Bazopoulou, N. Tavernarakis: The NemaGENETAG initiative: large scale transposon insertion gene-tagging in Caenorhabditis elegans. In: Genetica. Volume 137, Number 1, September 2009, pp. 39-46, ISSN  1573-6857 . doi : 10.1007 / s10709-009-9361-3 . PMID 19343510 .
  2. ↑ Y. Jiang, Z. Cai, W. Xie, T. Long, H. Yu, Q. Zhang: Rice functional genomics research: progress and implications for crop genetic improvement. In: Biotechnology Advances . Volume 30, Number 5, 2012 Sep-Oct, pp. 1059-1070, ISSN  1873-1899 . doi : 10.1016 / j.biotechadv.2011.08.013 . PMID 21888963 .
  3. ↑ M. Vandenbussche, J. Zethof, T. Gerats: Transposon display: a versatile method for transposon tagging. In: Methods in molecular biology (Clifton, NJ). Volume 1057, 2013, pp. 239-250, ISSN  1940-6029 . doi : 10.1007 / 978-1-62703-568-2_17 . PMID 23918433 .
  4. ↑ DG Moerman, RJ Barstead: Towards a mutation in every gene in Caenorhabditis elegans. In: Briefings in functional genomics & proteomics. Volume 7, Number 3, May 2008, pp. 195-204, ISSN  1477-4062 . doi : 10.1093 / bfgp / eln016 . PMID 18417533 . PDF .