Difference between revisions of "Part:BBa I714891"

Revision as of 06:38, 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.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 https://static.igem.org/mediawiki/parts/f/ff/GFPAFCM18.png
+<!-- Add more about the biology of this part here
+===Usage and Biology===
+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 PcaF).
+[[File:Scau-china-2018-11.png|800px|thumb|center|Figure 1 The result of the fluorescent intensity measurement ]]
+[[File:Scau-china-2018-12.png|800px|thumb|center|Figure 2 Fluorescent intensity of BBa_K2559006 driven by by PrplJ, Pdapa and PcaiF promoter.]]
+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.
+To expand the application of BBa_K2559005, we search the BBa_J04450 stored in Registry. 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.
 <!-- Add more about the biology of this part here