2014年11月
Gene Targeting by RNAi-Mediated Knockdown of Potent DNA Ligase IV Homologue in the Cellulase-Producing Fungus Talaromyces cellulolyticus
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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- 巻
- 174
- 号
- 5
- 開始ページ
- 1697
- 終了ページ
- 1704
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1007/s12010-014-1142-5
- 出版者・発行元
- HUMANA PRESS INC
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.
- リンク情報
- ID情報
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- DOI : 10.1007/s12010-014-1142-5
- ISSN : 0273-2289
- eISSN : 1559-0291
- PubMed ID : 25161035
- Web of Science ID : WOS:000344339700001