TABLE 2.
Human-yeast UBC9 chimeras
| Plasmida | Descriptionb |
|---|---|
| YEpGAL1h123yUBC9 · L | A human-yeast UBC9 chimera, PCR amplified from YCpUBC9, was cotransformed into EKY3 cells with NotI-linearized YCpGAL1hUBC9 · U; sequencing confirmed the chimera junction at residue 123 in plasmids recovered from uracil prototrophs, and the h123yUBC9 chimera was then inserted into the BamHI-NotI sites of YEpGAL1 · L |
| YEpGAL1h143yUBC9 · L | As above, a h143yUBC9 chimera was generated by PCR; the h143y chimera in recombinant URA3 plasmids was confirmed by sequencing |
| YEpGAL1h123y143hUBC9 · L | As above, a human-yeast-human UBC9 chimera was generated by PCR, followed by cotransformation with NotI-linearized YEpGAL1h123yUBC9 · L; the h123y143h chimera in recombinant URA3 plasmids was confirmed by sequencing. |
| YEpGAL1y123hUBC9 · L | As above, a yeast-human UBC9 chimera, PCR amplified from YCpGAL1hUBC9 · U, was cotransformed with NotI-linearized YCpUBC9; sequencing of plasmids recovered from uracil prototrophs confirmed the y123h chimera, which was then excised and ligated into YEpGAL1 · L |
| YEpGAL1y143hUBC9 · L | As above, a y143hUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1h22yUBC9 · L | A human-yeast UBC9 chimera was PCR amplified and cotransformed into EKY3 cells with BamHI-linearized YCpGAL1yUBC9 · U; sequencing confirmed the chimera junction at residue 22 in plasmids recovered from uracil prototrophs, and the h22yUBC9 chimera was then inserted into the BamHI-NotI sites of YEpGAL1 · L |
| YEpGAL1y22hUBC9 · L | As above, a y22hUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1h38yUBC9 · L | As above, a h38yUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1y38hUBC9 · L | As above, a y38hUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1h42yUBC9 · L | As above, a h42yUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1y42hUBC9 · L | As above, a y42hUBC9 chimera was generated by PCR and confirmed by sequencing |
| YEpGAL1y22h123y143hUBC9 · L | A y22hUBC9 chimera was generated in YEpGAL1h123y143hUBC9 · L by PCR and confirmed by sequencing |
| YEpGAL1y38h123y143hUBC9 · L | A y38hUBC9 chimera was generated in YEpGAL1h123y143hUBC9 · L by PCR and confirmed by sequencing |
a
The corresponding YCp series of vectors were also generated in pRS415.
b
Primer sequences are available upon request.