Part:BBa_K3930019

Inducer rtTA for the TetO7 promoter Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Introduction

The rtTA advanced cassette is composed by a promoter HXT7, the gene rtTA which allows the induction of the promoter Teto7 upon binding with doxycycline, and the terminator GYP7. This part is related to the arleady used part (BBa_K2748001), but we add to it the terminator and the promoter. It must be used with the part sequence of Teto7 promoter (BBa_K3930014). The sequence comes from the publication (Garí et al. 1997)

Results

Production of β-caotene (BBa_K3930003)

The part (BBa_K3930002) was linearized and transformed into the S.cerevisiae LycoYeast strain. The production of β-carotene was measured by HPLC. Figure 1 shows the chromatogram of a induced strain with 10 μg.ml-1 of doxycycline.

Figure 1: Production of β-carotene upon doxycycline activation

β-caroteneis produced in vivo by our strain. On the right are presented the mass spectra that correspond between the standard and the observed peak

This promoter Teto7 isn't negatively regulated and is always active under those lab conditions.

References

1. Garí E, Piedrafita L, Aldea M, Herrero E. 1997. A set of vectors with a tetracycline-regulatable promoter system for modulated gene expression in Saccharomyces cerevisiae. Yeast. 13(9):837–848. doi:10.1002/(SICI)1097-0061(199707)13:9<837::AID-YEA145>3.0.CO;2-T.