<!DOCTYPE html>

Parts

TokyoTech 2018 iGEM Team Parts

Name	Type	Description	Design	Length(bp)
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.

Biosafety

As 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.