TokyoTech 2018 iGEM Team Parts
|BBa_K2729000||Coding||DENV1 C||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||300|
|BBa_K2729001||Coding||DENV2 C||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||300|
|BBa_K2729002||Coding||DENV3 C||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||300|
|BBa_K2729003||Coding||DENV4 C||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||300|
|BBa_K2729004||Coding||DENV3 E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||1479|
|BBa_K2729005||Coding||DENV4 E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||1485|
|BBa_K2729006||Coding||EGFP_FMDV2a||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||950|
|BBa_K2729007||Coding||DsRed_FMDV2a||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||908|
|BBa_K2729008||Coding||ZsYellow_FMDV2a||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||920|
|BBa_K2729009||Coding||AmCyan_FMDV2a||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||920|
|BBa_K2729010||Coding||DENV1 AncC_prM_E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||2028|
|BBa_K2729011||Coding||DENV2 AncC_prM_E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||2028|
|BBa_K2729012||Coding||DENV3 AncC_prM_E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||2022|
|BBa_K2729013||Coding||DENV4 AncC_prM_E||Hayato Ito, Kotaro Miyamoto, Hajime Fujita||2028|
Note: Validated Parts (Silver Medal Criteria)DENV2 C, EGFP-FMDV2a and DsRed-Express-FMDV2a are essential for pseudo-virus production. As one of the structural genes, DENV2 C plays an important role in virus structural formation. EGFP-FMDV2a and DsRed-Express-FMDV2a are enclosed in pseudo-virus and programmed to code functional fluorescence proteins after infecting the host cells. To get specific sequence and charanterization information, please see the following Parts Pages.
Figure: List of validated parts and the features
Best Parts: EGFP-FMDV2a (BBa_K2729006)
Our team made new pseudovirus that can infect Vero cells and introduce the gene including this EGFP-FMDV2a. With the assistance of "self-cleaving" 2A peptides, EGFP is produced inside of the cell and folded so that it can emit fluorescence at the end.
Since EGFP is 35 times brighter than wild-type GFP and FMDV2a precisely works in our assumption, you can easily check the degree of infection by measuring fluorescence intensity.
BiosafetyAs you can see in Fig. 1, on native virus genome, structural genes and non-structural genes are integrated on the same sequence. Thus, the whole genome are replicated and that make viruses possible to replicate themselves in host cells. However, as you can see our construct design in Fig. 2, structural and non-structural regions are splitted so that the pseudo-virus cannot replicate themselves in host cells because RNA polymerase from non-structural V gene cannot replicate structural genes. To sum up, our system starts and ends in one place and doesn't harm environment.
Figure 1: Infection process of native virus and production of infectious pseudo-virus
Figure 2: Infection process of our pseudo-virus for validating biosafety