Difference between revisions of "Part:BBa K2997000"
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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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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 278
- 1000COMPATIBLE WITH RFC[1000]