Difference between revisions of "Part:BBa K2997000"

(Expression in E. coli)
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We amplified the tyrP gene and its promoter from E. coli MG1655 and cloning into pSB4A3. Then transformed the plasmid into DH5a and E. coli Nissle 1917, and extracted the plasmid after the colony formed and did the enzyme digestion to confirm the insertion was successful.
 
We amplified the tyrP gene and its promoter from E. coli MG1655 and cloning into pSB4A3. Then transformed the plasmid into DH5a and E. coli Nissle 1917, and extracted the plasmid after the colony formed and did the enzyme digestion to confirm the insertion was successful.
 
[[File:T--NCKU Tainan--Result-TyrP-double digest 2.jpg]]
 
[[File:T--NCKU Tainan--Result-TyrP-double digest 2.jpg]]
 
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<img src="https://2019.igem.org/wiki/images/a/a7/T--NCKU_Tainan--Result-TyrP-double_digest_2.jpg">
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 13:56, 18 October 2019


Tyrosine transporter (tyrP)

Background

It’s the gene encode for tyrosine transporter that is involved in transporting tyrosine across the cytoplasmic membrane. Hence, we used PCR to amplify this gene from E. coli MG1655 and incorporated it to our E. coli, allowing it to take in more tyrosine.

Expression in E. coli

We amplified the tyrP gene and its promoter from E. coli MG1655 and cloning into pSB4A3. Then transformed the plasmid into DH5a and E. coli Nissle 1917, and extracted the plasmid after the colony formed and did the enzyme digestion to confirm the insertion was successful. File:T--NCKU Tainan--Result-TyrP-double digest 2.jpg Sequence and Features


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 AgeI site found at 278
  • 1000
    COMPATIBLE WITH RFC[1000]