Difference between revisions of "Part:BBa K2559005"

Revision as of 07:13, 16 October 2018

Amended eGFP

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 I714891) 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).

Figure 1 The result of the fluorescent intensity measurement

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.

Because the BBa_I714891 is not available, we chose the BBa_J04450stored in Registry and tried 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

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]

@@ Line 6: / Line 6: @@
 ===Usage and Biology===
-The part BBa_K2559006 has an improvement on the basic part submitted by iGEM07_Peking ([https://parts.igem.org/Part:BBa I714891# BBa I714891]) 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.
+The part BBa_K2559006 has an improvement on the basic part submitted by iGEM07_Peking ([https://parts.igem.org/Part:BBa_I714891# BBa I714891]) 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 ]]
@@ Line 12: / Line 12: @@
 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 [https://parts.igem.org/Part:BBa_J04450# 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 [https://parts.igem.org/Part:BBa_K2559009# BBa_K2559009].
+Because the BBa_I714891 is not available, we chose the [https://parts.igem.org/Part:BBa_J04450# BBa_J04450]stored in Registry and tried 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.