Difference between revisions of "Part:BBa K319041:Design"

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===Results===
 
===Results===
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After BioBricking the nat cassette, this part was cloned into our novel [https://parts.igem.org/wiki/index.php?title=Part:BBa_K319043 Ade4 targeting vecoter] using standard BioBrick Protocols and was transformed into ''S. cerevisiae'' YPH500 strain. The YPH500 strain has a deletion in its ade2 locus; therefore,
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After BioBricking the nat cassette, this part was cloned into our novel [https://parts.igem.org/wiki/index.php?title=Part:BBa_K319043 ADE4 targeting vecoter] using standard BioBrick Protocols and was transformed into ''S. cerevisiae'' YPH500 strain. The YPH500 strain has a deletion in its ade2 locus; therefore, the colonies of this strain are red in the absence of adenine. However, when the ADE4 gene is knocked out, when our vector recombines, white colonies are formed again (Refer to our [[http://2010.igem.org/Team:uOttawa/Project project page]]).
  
 
===Source===
 
===Source===

Revision as of 23:51, 27 October 2010

NatMX cassette


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 690
    Illegal NgoMIV site found at 779
    Illegal AgeI site found at 408
  • 1000
    COMPATIBLE WITH RFC[1000]


Backgournd

The nat gene was first isolated from Streptomyces noursei[1]. This gene provides the organism with resistance to the antibiotic nourseothricin. In 1999, Goldstein and McCusker produced a nat cassette based on the kanMX cassettes of plasmid pFA6 and showed that this new cassette confers resistance to the above mentioned drug in S. cerevisiae strain YAG44[2]. We have BioBricked the complete cassette (including the upstream TEF1 promoter and the downstream TEF1 terminator) and have shown that it has 100% selectivity in S. cerevisiae YPH500 strain.

Results

After BioBricking the nat cassette, this part was cloned into our novel ADE4 targeting vecoter using standard BioBrick Protocols and was transformed into S. cerevisiae YPH500 strain. The YPH500 strain has a deletion in its ade2 locus; therefore, the colonies of this strain are red in the absence of adenine. However, when the ADE4 gene is knocked out, when our vector recombines, white colonies are formed again (Refer to our http://2010.igem.org/Team:uOttawa/Project project page).

Source

n

References