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发布时间：2019-03-12

论文类型：期刊论文

发表时间：2018-06-01

发表刊物：BIOTECHNOLOGY LETTERS

收录刊物：SCIE、PubMed

卷号：40

期号：6

页面范围：933-940

ISSN号：0141-5492

关键字：FLP/FRT system; Homologous recombination; Inducible expression; Nuclear localization signal sequence; Rhodosporidium toruloides

摘要：Objectives To establish a recombinase flippase (FLP) and flippase recognition target (FRT) system-mediated protocol for post-integration excision of exogenous DNA fragments in the oleaginous yeast Rhodosporidium toruloides.
   Results Binary vectors were constructed to harbor FLP expressing cassette together with the hygromycin-resistance marker. Results showed that R. toruloides transformants produced FLP, but failed to mediate removal of the bleomycin-resistance marker within two FRT sites. When FLP was fused with a native nuclear localization signal (NLS) peptide, the system was found functional. Moreover, R. toruloides recombinant strains expressing the NLS-fused FLP under the control of PADH2, an promoter of alcohol dehydrogenase 2 gene (RHTO_03062), were obtained to realize homologous recombination upon growing in glucose-deficient medium.
   Conclusions We have devised a homologous recombination method for R. toruloides based on the FLP/FRT system, which may facilitate further metabolic engineering and designing advanced cell factories for value-added chemicals.