Team:Tsinghua/Result3
From 2009.igem.org
GuoQiangChen (Talk | contribs) (→Targeted Biobrick Characterization by Fusing with GFP) |
GuoQiangChen (Talk | contribs) (→Targeted Biobrick Characterization by Fusing with GFP) |
||
Line 25: | Line 25: | ||
[[Image:transfection3.png|800px|center|thumb|Selected Cells for Specificity Testing before co-incubation. A.MRC-5 human diploid fibroblast cells. B.Hela cells. 10X10 magnification]] | [[Image:transfection3.png|800px|center|thumb|Selected Cells for Specificity Testing before co-incubation. A.MRC-5 human diploid fibroblast cells. B.Hela cells. 10X10 magnification]] | ||
- | [[Image:transfection4.png| | + | [[Image:transfection4.png|1500px|center|thumb|Specificity Testing after co-incubation with purified Targeted Biobrick. A.MRC-5, pEGFP+Targeted Biobrick, at bright field; B.MRC-5, pEGFP+Targeted Biobrick, under fluorescent microscopy; C.MRC-5, pEGFP empty vector control, at bright field; D. MRC-5, pEGFP empty vector control, under fluorescent microscopy; E.HeLa, pEGFP+Targeted Biobrick, at bright field; F.HeLa, pEGFP+Targeted Biobrick, under fluorescent microscopy; G.HeLa, pEGFP empty vector control, at bright field; H.HeLa, pEGFP empty vector control, under fluorescent microscopy. 10X10 magnification]] |
Revision as of 15:27, 19 October 2009
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.