Part:BBa_K2984022

PsaD Promoter A1-A3; High-level expression of genes

This part is a part of the Chlamy-HUB-Collection. For the effective and regulated expression of genes a promoter is required. The promoter is located at the 5’ region of the sense strand of the DNA and provides a binding site for the RNA-polymerase and transcription factors. In the nucleus of the green alga Chlamydomonas reinhardtii a gene encoding a chloroplast protein - PsaD - is located. Fischer and Rochaix (2001) showed that the ORF of this gene does not contain any introns and that the promoter drives a strong constitutive expression of the gene. Using the identified sequence of the promoter a vector for the high-level expression of endogenous and exogenous genes was created (Fischer and Rochaix, 2001).

This part was designed to be used with the MoClo standard and has A1-A3 overhangs.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 18
  Illegal XhoI site found at 842
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

For the effective and regulated expression of genes a promoter is required. The promoter is located at the 5’ region of the sense strand of the DNA and provides a binding site for the RNA-polymerase and transcription factors. In the nucleus of the green alga Chlamydomonas reinhardtii a gene encoding a chloroplast protein - PsaD - is located. Fischer and Rochaix (2001) showed that the ORF of this gene does not contain any introns and that the promoter drives a strong constitutive expression of the gene. Using the identified sequence of the promoter a vector for the high-level expression of endogenous and exogenous genes was created (Fischer and Rochaix, 2001).

Characterization

Fig.1 - Successful transformation of L0-PsaD_A1-A3 into E. coli.

References

1. Fischer, N., & Rochaix, J. D. (2001). The flanking regions of PsaD drive efficient gene expression in the nucleus of the green alga Chlamydomonas reinhardtii. Molecular Genetics and Genomics, 265(5), 888-894.