Difference between revisions of "Part:BBa K185033"
m
 
(2 intermediate revisions by the same user not shown)
Line 11: Line 11:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K185033 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K185033 SequenceAndFeatures</partinfo>
 +
 +
 +
==Characterization of an inverter of lac system([https://parts.igem.org/wiki/index.php?title=Part:BBa_K185033 BBa_K185033])==
 +
 +
It is an inverter which its downsteam gene will always be repressed by lacI protein unless IPTG is added. And the synthetic rate of the downsteam gene is low because the low binding efficiency of B0031.
 +
 +
;And I choose to assemble [https://parts.igem.org/wiki/index.php?title=Part:BBa_I712019 BBa_I712019] with it. Here is the data:
 +
:{| class="prettytable" style="border:1px solid"
 +
! Sample
 +
! OD
 +
! RLUs1
 +
! RLUs1/OD
 +
|-
 +
| A1
 +
| 2.64
 +
| 8090131
 +
| 3064444
 +
|-
 +
| A2
 +
| 1.83
 +
| 12033165
 +
| 6575500
 +
|-
 +
| B1
 +
| 2.74
 +
| 14369947
 +
| 5244506
 +
|-
 +
| B2
 +
| 1.67
 +
| 8374294
 +
| 5014547
 +
|-
 +
| C1
 +
| 1.79
 +
| 34906168
 +
| 19500653
 +
|-
 +
| C2
 +
| 1.58
 +
| 44358500
 +
| 28075000
 +
|-
 +
| D1
 +
| 0.7
 +
| 59173888
 +
| 84534126
 +
|-
 +
| D2
 +
| 0.89
 +
| 26208740
 +
| 29448022
 +
|-
 +
| E
 +
| 0.54
 +
| 26208740
 +
| 48534704
 +
|-
 +
| F
 +
| 1.45
 +
| 383217
 +
| 264287.6
 +
|}
 +
 +
#Sample A1/A2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 5h.
 +
#Sample B1/B2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 4h.
 +
#Sample C1/C2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 2h.
 +
#Sample D1/D2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 1h.
 +
#Sample E is BL21 strain carrying BBa_K185033, induced by IPTG. for 5min.
 +
#Sample F is BL21 strain carrying BBa_K185033, but it is not induced by IPTG.
 +
 +
 +
[[Image:SJTU09_Result_Overview6.jpg|center|thumb|750px|Fig 5. Relationship between inducing time(t) and RLUs1/OD ]]
 +
 +
From the data above, we know that the downstream genes (luciferase) are switched on sharply (about 100 times the uninduced sample) in the initial 1 hour. And then the synthesis of luciferase decreases into about 10 times the uninduced sample. It is certificated that our lac system is working.
  
  

Latest revision as of 12:26, 1 November 2009

An inverter of a special lactose operon system based on J23110-rbs34-lacI-dter-plac-rbs31

It is an inverter which its downsteam gene will always be repressed by lacI protein unless IPTG is added. And the synthetic rate of the downsteam gene is low because the low binding efficiency of B0031. This part improve on the basis of lac QPI system(BBa_K098986)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization of an inverter of lac system(BBa_K185033)

It is an inverter which its downsteam gene will always be repressed by lacI protein unless IPTG is added. And the synthetic rate of the downsteam gene is low because the low binding efficiency of B0031.

And I choose to assemble BBa_I712019 with it. Here is the data
Sample OD RLUs1 RLUs1/OD
A1 2.64 8090131 3064444
A2 1.83 12033165 6575500
B1 2.74 14369947 5244506
B2 1.67 8374294 5014547
C1 1.79 34906168 19500653
C2 1.58 44358500 28075000
D1 0.7 59173888 84534126
D2 0.89 26208740 29448022
E 0.54 26208740 48534704
F 1.45 383217 264287.6
  1. Sample A1/A2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 5h.
  2. Sample B1/B2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 4h.
  3. Sample C1/C2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 2h.
  4. Sample D1/D2 are BL21 strain carrying BBa_K185033, induced by IPTG. for 1h.
  5. Sample E is BL21 strain carrying BBa_K185033, induced by IPTG. for 5min.
  6. Sample F is BL21 strain carrying BBa_K185033, but it is not induced by IPTG.


Fig 5. Relationship between inducing time(t) and RLUs1/OD

From the data above, we know that the downstream genes (luciferase) are switched on sharply (about 100 times the uninduced sample) in the initial 1 hour. And then the synthesis of luciferase decreases into about 10 times the uninduced sample. It is certificated that our lac system is working.