Part:BBa_K555009
pYES2 - galactose-inducible expression vector for yeast
Galactose-inducible pYES2 vector for S. cerevisiae. Includes a multiple cloning site for gene insertion. See manufacturer's website for further information (http://products.invitrogen.com/ivgn/product/V82520?CID=AFLBC-Invitrogen). Note that this vector is sold by Invitrogen.
Plasmid map and growth curve
To get an overview of the growth of S. cerevisiae CENPK2-1D cells we compared the growth of the yeast cells carrying the empty vector in the not induced and the induced conditions. We used the growth profiler GP960 (EnzyScreen, Heemstede, The Netherlands), which is a temperature controlled shaker for microtiterplates with a scanner for growth detection. We incubated in square 24-deepwell plates with 1 ml of culture per well with a start OD of 1.
To get comparable data we adjusted all the start green values to 0, as they had the same OD and made a mean of the multiple parallel measurements to minimize the deviation errors.
The pyes2 vector has a galactose inducible promoter. For induction we added 10% of the volume of culture of 20% galactose.
The logarithmic growth phase can be determined from 4 to 7 hours after induction.
Comparing the induced and not induced cultures, it can be seen that they grow much better with additional galactose as S. cerevisiae can metabolize it as a carbon source. In order to improve the significance of these measurements addition of glucose to the not induced cells can be an outlook.
Furthermore, the cells carrying the MFalpha+mCherry inserts (BBa_K2020026 and BBa_E2060) are not affected in their growth after induction as the blue and yellow curves show.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 550
Illegal XbaI site found at 595
Illegal SpeI site found at 525
Illegal SpeI site found at 5851
Illegal PstI site found at 555
Illegal PstI site found at 3632
Illegal PstI site found at 4120 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 550
Illegal NheI site found at 3839
Illegal SpeI site found at 525
Illegal SpeI site found at 5851
Illegal PstI site found at 555
Illegal PstI site found at 3632
Illegal PstI site found at 4120
Illegal NotI site found at 576 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 550
Illegal BamHI site found at 519
Illegal XhoI site found at 583 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 550
Illegal XbaI site found at 595
Illegal SpeI site found at 525
Illegal SpeI site found at 5851
Illegal PstI site found at 555
Illegal PstI site found at 3632
Illegal PstI site found at 4120 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 550
Illegal XbaI site found at 595
Illegal SpeI site found at 525
Illegal SpeI site found at 5851
Illegal PstI site found at 555
Illegal PstI site found at 3632
Illegal PstI site found at 4120
Illegal NgoMIV site found at 5709
Illegal AgeI site found at 71 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 1996
Illegal BsaI.rc site found at 2942
Illegal SapI site found at 913
Illegal SapI site found at 3094
| None |