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Team:Wageningen UR/Journal/week24
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= Week 24: 8 october - 14 october = | = Week 24: 8 october - 14 october = | ||
| + | '''CCMV''' | ||
| + | ---- | ||
| + | |||
| + | ''12 October'' | ||
| + | |||
| + | Transformed IPTG_CCMV_NEG into a BL21 strain. Transformed biobrick RFP in pSB1C3 and the one with IPTG + RBS in kanamycin backbone into DH5A.<br/><br/> | ||
| + | |||
| + | ''14 October'' | ||
| + | Inoculated 10 mL LB with a colony from each transformation plate. | ||
| + | <br/><br/> | ||
'''GFP modification''' | '''GFP modification''' | ||
| Line 9: | Line 19: | ||
''11 October'' | ''11 October'' | ||
| - | * grow our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG | + | * grow JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter)and induce expression with IPTG |
-> no conclusion could be made due to growth on the negative control | -> no conclusion could be made due to growth on the negative control | ||
* send samples of BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing to check if the last transformation from pJET into the iGEM backbones succeded | * send samples of BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing to check if the last transformation from pJET into the iGEM backbones succeded | ||
| + | -> sequencing results where never obtained - the samples did not arrive | ||
''12 October'' | ''12 October'' | ||
| - | * 2nd time growing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG | + | * 2nd time growing JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG |
-> again there is growth on the negative control | -> again there is growth on the negative control | ||
-> no green fluorescence could be seen | -> no green fluorescence could be seen | ||
| + | |||
| + | ---- | ||
| - | ''' | + | '''PLRV''' |
---- | ---- | ||
| - | '' | + | We started another production process to obtain PLRV VLPs. We considered that production of the CCMV VLP with the negative inside had worked well in the first batch which produced with the well acknowledged production strain BL-21, and not in the later batch from the JM-109 strain. Because our last attempt to produce PLRV VLPs was done in JM-109 cells as well, we figure it may be possible that the production strain was the problem. Therefore, we will try the same procedure again but now in BL-21 ''E. coli''. |
| - | + | We transformed electrocompetent BL-21 cells with Bba_K883401, which contains a PLRV Coat Protein encoding gene under the IPTG inducible promotor. | |
| - | + | ||
---- | ---- | ||
[[https://2012.igem.org/Team:Wageningen_UR/Journal/week23 previous week]] [[https://2012.igem.org/Team:Wageningen_UR/Journal/week25 next week]] | [[https://2012.igem.org/Team:Wageningen_UR/Journal/week23 previous week]] [[https://2012.igem.org/Team:Wageningen_UR/Journal/week25 next week]] | ||
Latest revision as of 14:58, 26 October 2012
Week 24: 8 october - 14 october
CCMV
12 October
Transformed IPTG_CCMV_NEG into a BL21 strain. Transformed biobrick RFP in pSB1C3 and the one with IPTG + RBS in kanamycin backbone into DH5A.
14 October
Inoculated 10 mL LB with a colony from each transformation plate.
GFP modification
11 October
- grow JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter)and induce expression with IPTG
-> no conclusion could be made due to growth on the negative control
- send samples of BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing to check if the last transformation from pJET into the iGEM backbones succeded
-> sequencing results where never obtained - the samples did not arrive
12 October
- 2nd time growing JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG
-> again there is growth on the negative control -> no green fluorescence could be seen
PLRV
We started another production process to obtain PLRV VLPs. We considered that production of the CCMV VLP with the negative inside had worked well in the first batch which produced with the well acknowledged production strain BL-21, and not in the later batch from the JM-109 strain. Because our last attempt to produce PLRV VLPs was done in JM-109 cells as well, we figure it may be possible that the production strain was the problem. Therefore, we will try the same procedure again but now in BL-21 E. coli.
We transformed electrocompetent BL-21 cells with Bba_K883401, which contains a PLRV Coat Protein encoding gene under the IPTG inducible promotor.
