Team:Tsinghua/Result3
From 2009.igem.org
GuoQiangChen (Talk | contribs) (→C-Targeted Biobrick Fusion) |
GuoQiangChen (Talk | contribs) (→C-Targeted Biobrick Fusion) |
||
Line 17: | Line 17: | ||
=C-Targeted Biobrick Fusion= | =C-Targeted Biobrick Fusion= | ||
[[Image:C-fiber.png|300px|center|thumb|Identification of the cloned pACYCDuet-1+lysis+C-Targeted Biobrick+D+E+FI+FII. C stands for the PCR identification of C domain of C-Targeted Biobrick fusion protein. TB stands for the Targeted Biobrick domain of C-Targeted Biobrick fusion protein. pAC(TB) stands for the agarose gel electrophoresis of pACYCDuet-1+lysis+C-Targeted Biobrick+D+E+FI+FII]] | [[Image:C-fiber.png|300px|center|thumb|Identification of the cloned pACYCDuet-1+lysis+C-Targeted Biobrick+D+E+FI+FII. C stands for the PCR identification of C domain of C-Targeted Biobrick fusion protein. TB stands for the Targeted Biobrick domain of C-Targeted Biobrick fusion protein. pAC(TB) stands for the agarose gel electrophoresis of pACYCDuet-1+lysis+C-Targeted Biobrick+D+E+FI+FII]] | ||
+ | |||
+ | The construction of C-Targeted Biobrick fusion protein is identified by PCR. The fusion protein was directly engineered to the synthesized pACYCDuet-1+lysis+C+D+E+FI+FII. Thus, the positive clone, co-tranformed with pET-28a+Nu1+A+W+B, will be able to produce the targeted GenSniper with the desired fiber of defined specificity. However, we chose to characterize the function of Targeted Biobrick firstly according to our synthetic biology background. When we confirm that the GenSniper viroin (encoded by its genome and with the Therapeutic DNA packaged as showed in Module I and II) and Targeted Biobrick (Module III) can work separately. Then the characterization of their co-function (all the three modules) will be more definitive. | ||
=Targeted Biobrick Characterization by Fusing with GFP= | =Targeted Biobrick Characterization by Fusing with GFP= |
Revision as of 16:06, 19 October 2009
Contents |
Targeted Biobrick Construction
In this section, we were contructing two contructs in paralell. First, to contruct a standardized Targeted Biobrick for parts registry, where Targeted Biobrick can be served for further use by other teams or researchers. Second, to incorporate Targeted Biobrick into an expression vector pET-28a to test its expression. Targeted Biobrick was contructed via two rounds of PCR.
The expression vector was used for Targeted Biobrick characterization, while the standardized Targeted Biobrick has been submitted to iGEM headquarters for parts registry. We have successfully gained the desired Targeted Biobrick protein of 44184.58 Daltons.
In conclusion, we have successfully contructed Targeted Biobrick for both part registry and expression (most of the results have not been showed here to make our wiki concise). Of course, the next step should be the characterization of its targeted function.
C-Targeted Biobrick Fusion
The construction of C-Targeted Biobrick fusion protein is identified by PCR. The fusion protein was directly engineered to the synthesized pACYCDuet-1+lysis+C+D+E+FI+FII. Thus, the positive clone, co-tranformed with pET-28a+Nu1+A+W+B, will be able to produce the targeted GenSniper with the desired fiber of defined specificity. However, we chose to characterize the function of Targeted Biobrick firstly according to our synthetic biology background. When we confirm that the GenSniper viroin (encoded by its genome and with the Therapeutic DNA packaged as showed in Module I and II) and Targeted Biobrick (Module III) can work separately. Then the characterization of their co-function (all the three modules) will be more definitive.