We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Developing a flippase-mediated maker recycling protocol for the oleaginous yeast Rhodosporidium toruloides.
- Authors
Sun, Wenyi; Yang, Xiaobing; Wang, Xueying; Jiao, Xiang; Zhang, Sufang; Luan, Yushi; Zhao, Zongbao K.
- Abstract
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.
- Subjects
RECOMBINASES; SIGNAL peptides; RECOMBINANT proteins; ALCOHOL dehydrogenase; PROTEIN expression
- Publication
Biotechnology Letters, 2018, Vol 40, Issue 6, p933
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-018-2542-3