Gene targeting by RNAi-mediated knockdown of potent DNA ligase IV homologue in the cellulase-producing fungus Talaromyces cellulolyticus.

• Hayata, Koutarou

• Asada, Seiya

• Fujii, Tatsuya

• Inoue, Hiroyuki

• Ishikawa, Kazuhiko

• 澤山, 茂樹

抄録

The genome of the cellulase-producing fungus Talaromyces cellulolyticus (formerly Acremonium cellulolyticus) was screened for a potent DNA ligase IV gene (ligD homologue). Homologous recombination efficiency in T. cellulolyticus is very low. Therefore, suppression of a non-homologous end-joining system was attempted to enable specific gene knockouts for molecular breeding. The transcript levels of ligD homologue were 0.037 of those of the parental YP-4 strain in the Li20 transformant carrying the RNAi construct targeting the ligD homologue. Transformation of the hairpin-type RNAi vector into T. cellulolyticus could be useful in fungal gene knockdown experiments. Cellulase production and protein secretion were similar in the parental YP-4 strain and the Li20 transformant. Knockout transformation of ligD homologue using the Li20 transformant led to 23.1 % double crossover gene targeting. Our results suggest that the potent DNA ligase IV gene of T. cellulolyticus is related to non-homologous end joining and that the knockdown of the ligD homologue is useful in gene targeting.

収録刊行物

• Applied biochemistry and biotechnology

  Applied biochemistry and biotechnology 174 (5), 1697-1704, 2014-11

  Springer US

キーワード

• Homologous recombination
• DNA ligase IV
• RNAi
• Acremonium cellulolyticus
• Cellulose/biosynthesis
• Cellulose/genetics
• DNA Ligases/genetics
• Gene Knockdown Techniques/methods
• Gene Targeting
• Genetic Enhancement/methods
• RNA Interference/physiology
• Recombination, Genetic/genetics
• Talaromyces/genetics

詳細情報

• CRID: 1050001335813589248
• NII論文ID: 120005649316
• NII書誌ID: AA10621790
• ISSN: 02732289
• HANDLE: 2433/199675
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • CiNii Articles