Difference between revisions of "Part:BBa I714891"
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Because the BBa_I714891 is not avalible, we search the BBa_J04450 stored in Registry and to expand the application of BBa_K2559005.The BBa_J04450 part is a strong RFP expression vector for E.coli because of the strong RBS. As the main page BBa_J04450 mentioned, the E.coli colonies with BBa_J04450 were red in color under natural light after about 18 hour culture in LB plate. We used the BBa_K2559005 to replace the RFP region in BBa_J04450, the restructured vector from those two parts is [https://parts.igem.org/Part:BBa_K2559009# BBa_K2559009]. | Because the BBa_I714891 is not avalible, we search the BBa_J04450 stored in Registry and to expand the application of BBa_K2559005.The BBa_J04450 part is a strong RFP expression vector for E.coli because of the strong RBS. As the main page BBa_J04450 mentioned, the E.coli colonies with BBa_J04450 were red in color under natural light after about 18 hour culture in LB plate. We used the BBa_K2559005 to replace the RFP region in BBa_J04450, the restructured vector from those two parts is [https://parts.igem.org/Part:BBa_K2559009# BBa_K2559009]. | ||
We transferred the BBa_K2559009 to DH5α by heat-shock transformation, and found that the fluorescence signal can be observe clearly under the UV. | We transferred the BBa_K2559009 to DH5α by heat-shock transformation, and found that the fluorescence signal can be observe clearly under the UV. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 06:57, 16 October 2018
SDY_EGFP
a strong reporter
measured by fluorescence microscope
[http://2018.igem.org/Team:AFCM-Egypt# Team:AFCM-Egypt 2018] provided specific characterization for this part by using eGFP for experimental characterization of lentiviral transfer plasmid transfection efficacy and apoptotic effect on colorectal cancer cell line RKO.
The BBa_K2559005 is a full-length EGFP coding part improved from BBa_l714891.
Usage and Biology
The part BBa_K2559006 has an improvement on the basic part submitted by iGEM07_Peking (BBa_l714891) which is cording the SDY_EGFP. We had added 16 bp DNA to fill the length of EGFP cording sequence in BBa_l714891 to crease the BBa_K2559005. Because, we found out a 16 bp deficiency in the EGFP starting cording region of BBa_l714891, after we checked the sequence of BBa_l714891 in NCBI.
In order to test the function of BBa_K2559005, we design a new E.coli expression vector with our new part, BBa_K2559003, a strong E.coli endogenous promoter (PrplJ). The full length EGFP in BBa_K2559005 was driven by PrplJ, and expressed in DH10B. Attention, we also used the BBa_K2559005 in the promoter strength analysis of our other two new parts, the BBa_K2559004 and BBa_K2559011 which are weak E.coli endogenous promoters (PdapA and PcaiF).
We conclude that our improved new part, full length EGFP cording biobrick , BBa_K2559005 can work well in DH10B, and we also hoped that our improvement on the BBa_l714891 can help the future application of this part.
Because the BBa_I714891 is not avalible, we search the BBa_J04450 stored in Registry and to expand the application of BBa_K2559005.The BBa_J04450 part is a strong RFP expression vector for E.coli because of the strong RBS. As the main page BBa_J04450 mentioned, the E.coli colonies with BBa_J04450 were red in color under natural light after about 18 hour culture in LB plate. We used the BBa_K2559005 to replace the RFP region in BBa_J04450, the restructured vector from those two parts is BBa_K2559009. We transferred the BBa_K2559009 to DH5α by heat-shock transformation, and found that the fluorescence signal can be observe clearly under the UV.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]