Difference between revisions of "Part:BBa K2559005"

(→‎Usage and Biology)
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===Usage and Biology===
 
===Usage and Biology===
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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.
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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).
 
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-2018-11.png|800px|thumb|center|Figure 1 The result of the fluorescent intensity measurement ]]
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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.
 
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.
  
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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.  
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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].  
 
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.  
  

Revision as of 07:08, 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 (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).

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.

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.


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]