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Team:Lethbridge Canada/Results
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'''RFP with Cells Media at Different Induction Levels''' | '''RFP with Cells Media at Different Induction Levels''' | ||
| + | Preliminary results | ||
Cultures were grown containing pLacI promoter attached to a Twin Arginine tag signal sequence and fused to red fluorescent protein. Samples were taken at various time points and examined for fluorescents when excited at 586nm with an expected emission at 607nm. We used a fluorescents spectrophotometer to measure the red fluorescent protein present in the supernatant after spinning the cells to a pellet. We used a constutively expressed construct expressing Red fluorescent protein( J23100 in J61002) | Cultures were grown containing pLacI promoter attached to a Twin Arginine tag signal sequence and fused to red fluorescent protein. Samples were taken at various time points and examined for fluorescents when excited at 586nm with an expected emission at 607nm. We used a fluorescents spectrophotometer to measure the red fluorescent protein present in the supernatant after spinning the cells to a pellet. We used a constutively expressed construct expressing Red fluorescent protein( J23100 in J61002) | ||
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| - | [[Image:Time A.JPG|left|px]][[Image:Time B.JPG|right|px]] | + | [[Image:Time A.JPG|left|px]] |
| + | Fluorescents was measured one hour after induction, no difference could be seen between the test and control constructs in the supernatant . | ||
| + | [[Image:Time B.JPG|right|px]] | ||
| + | Fluorescents was measured two hours after induction, no difference could be seen between the test and control constructs in the supernatant . | ||
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| - | [[Image:Time C.JPG|left|px]][[Image:Time D.JPG|right|px]] | + | [[Image:Time C.JPG|left|px]] |
| + | Fluorescents was measured three hours after induction, no difference could be seen between the test and control constructs in the supernatant . | ||
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| + | [[Image:Time D.JPG|right|px]] | ||
| + | Fluorescents was measured four hours after induction, no difference could be seen between the test and control constructs in the supernatant . | ||
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Revision as of 06:27, 17 June 2012
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ResultsParts createdWe created two new parts, [http://partsregistry.org/Part:BBa_K736000 BBa_K736000] (which regulated the transcription of downstream genes) and [http://partsregistry.org/Part:BBa_K736001 BBa_K736001] (which was a TAT signal sequence fused to a red fluorescent protein, E1010). We also assembled two parts, [http://partsregistry.org/wiki/index.php?title=Part:BBa_K736002 BBa_K736002] (which tests the ability of glucose to regulate transcription and uses enhanced RFP as the reporter) and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K736003 BBa_K736003] (which tests for the ability of the TAT sequence to cause secretion of protein outside of the cell and also uses RFP as a reporter). We also created one part that coded for human insulin chain A ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K736004 BBa_K736004]) and one part that coded for human insulin chain B ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K736005 BBa_K736005]). We are also submitting the DNA for [http://partsregistry.org/Part:BBa_K331009 K331009] which was in its planning stage in the registry.
Proof of PrincipleTAT signal sequence RFP with Cells Media at Different Induction Levels Preliminary results Cultures were grown containing pLacI promoter attached to a Twin Arginine tag signal sequence and fused to red fluorescent protein. Samples were taken at various time points and examined for fluorescents when excited at 586nm with an expected emission at 607nm. We used a fluorescents spectrophotometer to measure the red fluorescent protein present in the supernatant after spinning the cells to a pellet. We used a constutively expressed construct expressing Red fluorescent protein( J23100 in J61002)
 Fluorescents was measured one hour after induction, no difference could be seen between the test and control constructs in the supernatant .  Fluorescents was measured two hours after induction, no difference could be seen between the test and control constructs in the supernatant .
 Fluorescents was measured three hours after induction, no difference could be seen between the test and control constructs in the supernatant .  Fluorescents was measured four hours after induction, no difference could be seen between the test and control constructs in the supernatant .
  
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