Back to article: Yeast for virus research

FIGURE 2: Schematic diagram of a shotgun approach to clone a small viral genome in fission yeast. (Top) the pYZ1N vector contains a wild type nmt1 promoter [68, 196]. It carries a LEU2 gene for selection. pYZ2N contains the wild type nmt1 promoter and a URA4 gene selection marker. (Bottom) pYZ3N-GFP contains the same nmt1 promoter as pYZ1N and has an added green fluorescent protein GFP tag. The α-peptide of β-galactosidase is used for selecting DNA inserts with α-complementation. Unique cloning sites in these vectors are indicated. ars1, origin of replication from S. pombe; Leu2, Saccharomyces cerevisiae leucine biosynthesis gene; AmpR, bacterial ampicillin resistance gene. Adapted from [36, 68]. Note that nothing is new in molecular features of shuttle vectors described. Goal of this figure is to illustrate a robust and streamlined strategy of shotgun gene cloning of a small viral genome.

36. Li G, Poulsen M, Fenyvuesvolgyi C, Yashiroda Y, Yoshida M, Simard JM, Gallo RC, Zhao RY (2017). Characterization of cytopathic factors through genome-wide analysis of the Zika viral proteins in fission yeast. Proc Natl Acad Sci U S A 114(3): E376-E385. 10.1073/pnas.1619735114
68. Zhao Y, Elder RT, Chen M, Cao J (1998). Fission yeast expression vectors adapted for positive identification of gene insertion and green fluorescent protein fusion. Biotechniques 25(3): 438-440, 442, 444.
196. Maundrell K (1990). nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine. J Biol Chem 265(19): 10857-10864.

By continuing to use the site, you agree to the use of cookies. more information

The cookie settings on this website are set to "allow cookies" to give you the best browsing experience possible. If you continue to use this website without changing your cookie settings or you click "Accept" below then you are consenting to this. Please refer to our "privacy statement" and our "terms of use" for further information.