Difference between revisions of "Part:BBa K567016"
 
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<partinfo>BBa_K567016 short</partinfo>
 
<partinfo>BBa_K567016 short</partinfo>
  
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This biobrick is constructed by mutating the anticodon of tRNA&#65288;met&#65289; to TCG (base pairing codon CGA). This tRNA can transfer fMet to CGA when it is used as the start codon. Kana gene with start codon substituted for CGA is used to testify the function of metY-CGA. When this biobrick and metGM(BBa_K567014) or metG(BBa_K567015) are co-transformed into the cell, the cells can survive on the LBKana plate.  
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This biobrick is constructed by mutating the anticodon of tRNA<sup>Met</sup> to TCG (base pairing codon CGA). This tRNA can transfer fMet to CGA when it is used as the start codon. KanR gene with start codon substituted for CGA is used to testify the function of ''metY''-CGA. When this biobrick and ''metG''M(BBa_K567014) or ''metG''N(BBa_K567015) are co-transformed into the cell, the cells can survive on the LB Kan plate.  
  
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
 
===Construction of BBa_K567016===
 
===Construction of BBa_K567016===
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===Characterization of BBa_K567016===
 
===Characterization of BBa_K567016===
  
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When this part, MetRS PT7-''metG''N (BBa_K567015)(or MetRS PT7-''metG''M (BBa_K567014)) and KanaR (BBa_K567020) are co-transformed into the cell, the cell is expected to survive Kana.  
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When this part, MetRS PT7-''metG''N (BBa_K567015)(or MetRS PT7-''metG''M (BBa_K567014)) and KanR (BBa_K567020) are co-transformed into the cell, the cell is expected to survive Kanamycin.  
  
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[[image:11SJTU-initial_codon_result.jpg|frame|center|fig. Growth of ER2566 with a. ''metG''N + ''metY''-CGA, b. ''metG''M + ''metY''-CGA, c. + ''metG''N, d. + ''metG''M. Growth medium (left): LB Kana+Tet. Growth medium (right): LB Kana.]]
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[[image:11SJTU-initial_codon_result.jpg|frame|center|Fig. Growth of ER2566 with a. ''metG''N + ''metY''-CGA, b. ''metG''M + ''metY''-CGA, c. + ''metG''N, d. + ''metG''M. Growth medium (left): LB Kana+Tet. Growth medium (right): LB Kana.]]
  
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Cell growth shows that the cells show Kana resistance only when both modified MetRS (''metG''N) and modified tRNA<sup>Met</sup>(''metY''-CGA) are transformed into the cell, proving that ''metG''N works well.
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Cell growth shows that the cells show Kanamycin resistance only when both modified MetRS (''metG''N) and modified tRNA<sup>Met</sup>(''metY''-CGA) are transformed into the cell, proving that ''metG''N works well.
  
  
 
For more information concerning this part, please see [http://2011.igem.org/Team:SJTU-BioX-Shanghai 2011 SJTU-BioX-iGEM]
 
For more information concerning this part, please see [http://2011.igem.org/Team:SJTU-BioX-Shanghai 2011 SJTU-BioX-iGEM]
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Related Biobrick:
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PT7-''metG''M  ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567014 BBa_K567014])
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PT7-''metG''N  ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K567015 BBa_K567015])
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:32, 29 October 2011

metY-CGA

This biobrick is constructed by mutating the anticodon of tRNAMet to TCG (base pairing codon CGA). This tRNA can transfer fMet to CGA when it is used as the start codon. KanR gene with start codon substituted for CGA is used to testify the function of metY-CGA. When this biobrick and metGM(BBa_K567014) or metGN(BBa_K567015) are co-transformed into the cell, the cells can survive on the LB Kan plate.


Construction of BBa_K567016

metY-CGA: We have cloned operon metY containing tRNAMet from E.coli to pACYC184. The anticodon of tRNAMet was mutated to TCG (base pairing codon CGA).


Characterization of BBa_K567016

When this part, MetRS PT7-metGN (BBa_K567015)(or MetRS PT7-metGM (BBa_K567014)) and KanR (BBa_K567020) are co-transformed into the cell, the cell is expected to survive Kanamycin.

Fig. Growth of ER2566 with a. metGN + metY-CGA, b. metGM + metY-CGA, c. + metGN, d. + metGM. Growth medium (left): LB Kana+Tet. Growth medium (right): LB Kana.

Cell growth shows that the cells show Kanamycin resistance only when both modified MetRS (metGN) and modified tRNAMet(metY-CGA) are transformed into the cell, proving that metGN works well.


For more information concerning this part, please see [http://2011.igem.org/Team:SJTU-BioX-Shanghai 2011 SJTU-BioX-iGEM]

Related Biobrick:

PT7-metGM (BBa_K567014)

PT7-metGN (BBa_K567015)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 183
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 183
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 183
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 183
  • 1000
    COMPATIBLE WITH RFC[1000]