Active human full-length CDKL5 produced in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125

Table 1 Characteristics of BCD constructs for flCDKL5 expression

Name	Leader^a	SD1^b	SD2^c	N-tags^d	C-tags^d	Mutations^d
p79C-107 (G)	/	/	CAACAGGAA	TATκ	3xFLAG-6xHis	/
pBCD1-107 (G)	LacZ	CAACAGGAA	CAACAGGAA	TATκ	3xFLAG-6xHis	/
pBCD2-107 (G)	LacZ	CAACAGGAA	AAGGAGGTC	TATκ	3xFLAG-6xHis	/
pBCD1-107 (K)	LacZ	CAACAGGAA	CAACAGGAA	6xHis-TATκ	3xFLAG	/
pBCD2-107 (K)	LacZ	CAACAGGAA	AAGGAGGTC	6xHis-TATκ	3xFLAG	/
pBCD3-107 (K)	TrpA	AAGGAGGTC	AAGGAGGTC	6xHis-TATκ	3xFLAG	/
pBCD2-107 (G) M10V	LacZ	CAACAGGAA	AAGGAGGTC	TATκ	3xFLAG-6xHis	M43V
pBCD2-107 (G) STOP	LacZ	CAACAGGAA	AAGGAGGTC	TATκ	3xFLAG-6xHis	M1I, G2STOP
pBCD2-107 (K) M10V	LacZ	CAACAGGAA	AAGGAGGTC	6xHis-TATκ	3xFLAG	M61V
pBCD2-107 (L) M10V	LacZ	CAACAGGAA	AAGGAGGTC	6xHis-Sumo-TATκ	3xFLAG	M159V

^aLeader peptide encoded upstream of flCDKL5 includes the firs 19 residues of the indicated proteins. ^bShine Dalgarno sequence upstream of the Leader peptide encoding sequence. ^cShine Dalgarno sequence upstream of the flCDKL5 gene. Shine Dalgarno sequences which are entirely complementary to 16S rRNA are underlined. ^dN- and C-terminal tags and mutations are referred to flCDKL5 protein. M43V, M61V and M159V are the same mutation (M10V in human CDKL5, O76039, Uniprot), but the coordinates differ because of the presence of different N-terminal tags

ISSN: 1475-2859