Difference between revisions of "Part:BBa K2984022"
 
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<partinfo>BBa_K2984022 parameters</partinfo>
 
<partinfo>BBa_K2984022 parameters</partinfo>
 
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==Characterization==
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<img src="https://static.igem.org/mediawiki/parts/f/f2/T--Humboldt_Berlin--PsaD_A1-A3.png" alt="Plate_L0-PsaD_A1-A3_E.coli" width="500">
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<figcaption>Fig.1 - Successful transformation of L0-PsaD_A1-A3 into <i>E. coli</i>.</figcaption>
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===Functional Parameters===
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<partinfo>BBa_K2984009 parameters</partinfo>
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Latest revision as of 13:14, 20 October 2019


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

Plate_L0-PsaD_A1-A3_E.coli
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.