Creation of universal vector collections for efficient epitope tagging and gene deletion in filamentous fungi

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	Creation of universal vector collections for efficient epitope tagging and gene deletion in filamentous fungi
	Qin, L.; Tan, K.; Li, A.; Guo, S.; Chen, Y.; Wang, F.; Wong, K. H.
	2017-08-22
Source Publication	XII International Fungal Biology Conference
Abstract	In the post-genomic era, gene manipulation (e.g. deletion, epitope-tagging, over-expression, etc.) via homologous recombination has become a routine for validating observations and testing hypotheses gained from high-throughput Genomics and Bioinformatics studies. Unlike Saccharomyces cerevisiae, homologous recombination in filamentous fungi requires long homologous flanking arms, and generation of transformation constructs is often one of the rate limiting steps in gene manipulation experiments. In this study, we have modified and expanded existing S. cerevisiae vector collections (Longtine et al., 1998 Yeast 14(10):953-61 and Funakoshi and Hochstrasser Yeast 26(3):185-192) to be used for gene deletion and epitope-tagging in filamentous fungi. For the gene deletion vector collection, three commonly used auxotrophic markers (Af-pyrG, Af-riboB and Af-pyroA) and a dominant glufosinate resistant marker are added to the original collection carrying different drug resistant genes (Hygromycin, G418 and CloNAT). On the other hand, the vector collection for C-terminal epitope-tagging contains 40 vectors covering all combinations of 10 different epitope tags with the 4 selectable markers mentioned above. Using these vectors, we have also optimized a simple and efficient system for making deletion and tagging constructs for transformation. This system offers a quick way to swap epitope tags when an introduced epitope affects the function of a tagged protein or when a different epitope is desired. It is expected that the both vector collections will facilitate gene manipulation chores in filamentous fungi.
Keyword	Universal vector filamentous fungi
Language	英語English
The Source to Article	PB_Publication
PUB ID	32145
Document Type	Conference paper
Collection	DEPARTMENT OF BIOMEDICAL SCIENCES Faculty of Health Sciences
Corresponding Author	Wong, K. H.
Recommended Citation GB/T 7714	Qin, L.,Tan, K.,Li, A.,et al. Creation of universal vector collections for efficient epitope tagging and gene deletion in filamentous fungi[C], 2017.
APA	Qin, L.., Tan, K.., Li, A.., Guo, S.., Chen, Y.., Wang, F.., & Wong, K. H. (2017). Creation of universal vector collections for efficient epitope tagging and gene deletion in filamentous fungi. XII International Fungal Biology Conference.

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