Difference between revisions of "Part:BBa K594009:Design"

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===Source===
 
===Source===
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We got the Rhizobium etli CFN42 from Nanjing Agriculture University, and then cloned the raiI gene from Rhizobium etli CFN42 with primers designed by the part design tool in the website [ginkgobioworks.com]. We get its gene sequence from the www.microbesonline.org, and then cloned them through PCR with biobrick part primers.
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We got the Rhizobium etli CFN42 from Nanjing Agriculture University, but we fail to get the sequence of praiI, so we deduct a 500bp sequence that may cover the promoter. then cloned the deducted praiI gene from Rhizobium etli CFN42 with primers designed by the part design tool in the website [ginkgobioworks.com].
  
 
===References===
 
===References===

Latest revision as of 00:37, 6 October 2011

the promoter of raiI


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 444
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 447


Design Notes

The Rhizobium etli CFN42 is also a mode organism in Rhizobia. We looked for the papers related to the Quorum Sensing systems in Rhizobium etli CFN42, and then found the specific sequence in NCBI and asked the Rhizobium etli CFN42 from Nanjing Agricultural University. The raiI/R system have no threats to biosafety.

Source

We got the Rhizobium etli CFN42 from Nanjing Agriculture University, but we fail to get the sequence of praiI, so we deduct a 500bp sequence that may cover the promoter. then cloned the deducted praiI gene from Rhizobium etli CFN42 with primers designed by the part design tool in the website [ginkgobioworks.com].

References

1.VIOLA ROSEMEYER et al.luxI- and luxR-Homologous Genes of Rhizobium etli CNPAF512 Contribute to Synthesis of Autoinducer Molecules and Nodulation of Phaseolus vulgaris.JOURNAL OF BACTERIOLOGY,Feb. 1998, p. 815–821.
2.Anne Edwards,1 Marijke Frederix,1 Florence Wisniewski-Dye´,2 Jacob Jones,1 Angeles Zorreguieta,3 and J. Allan Downie1*.The cin and rai Quorum-Sensing Regulatory Systems in Rhizobium leguminosarum Are Coordinated by ExpR and CinS, a Small Regulatory Protein Coexpressed with CinI.JOURNAL OF BACTERIOLOGY, May 2009, p. 3059–3067
3.Juan E. Gonza´lez and Melanie M. Marketon. Quorum Sensing in Nitrogen-Fixing Rhizobia, MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 2003,p. 574–592