Key roles of amylopectin synthesis and degradation enzymes in the establishment and reactivation of chronic toxoplasmosis

Chen, Pu; Lyu, Congcong; Wang, Yidan; Pan, Ming; Lin, Xingyu; Shen, Bang

doi:10.1186/s44149-023-00083-x

Table 1 Primers used in this study

From: Key roles of amylopectin synthesis and degradation enzymes in the establishment and reactivation of chronic toxoplasmosis

Name	Sequence (5′ → 3′)	Used for
gRNA-USP-Fw	GTCTGTGTCACCTCCGTTCTGTTTTAGAGCTAGAAATAGC	To construct the USP-specific CRISPR plasmid
gRNA-GWD-Fw	GCCTGTGCACCAACTTCTTC GTTTTAGAGCTAGAAATAGC	To construct the GWD-specific CRISPR plasmid
gRNA-Rv	AACTTGACATCCCCATTTAC	To construct gene-specific CRISPR plasmids
DHRR-Fw	CAGGCTGTAAATCCCGTGAG	Amplification of DHFR fragment
DHFR-Rv	GATTCCGTCAGCGGTCTGTC	Amplification of DHFR fragment
U5-USP-Fw	GTGAGGAAACAAGTGCC	Amplification of 5′-homology of USP for 5H-DHFR*-3H construction
U5-USP-Rv	CTCACGGGATTTACAGCCTG CGCGTTTCCTTCAGCTT
U3-USP-Fw	GACAGACCGCTGACGGAATC TTTCGGAGTTCAAGTTGC	Amplification of 3′-homology of USP for 5H-DHFR*-3H construction
U3-USP-Rv	AGGCGAACCTCCACTT
U5-GWD-Fw	CGACGGCCAGTGAATTCGAG	Amplification of 5′-homology of GWD for 5H-DHFR*-3H construction
U5-GWD-Rv	GATGATTTGTGAGGACGACTCACGGGATTTACAGCCTGTC
U3-GWD-Fw	ACAGACCGCTGACGGAATCGGTGCGAGTCAAGAACTCAGG	Amplification of 3′-homology of GWD for 5H-DHFR*-3H construction
U3-GWD-Rv	GCTATGACCATGATTACGCCATCAG
5′-UpU5USP	GCTCGACATCTCTTCTCGGAG	PCR1 of Δusp
5′-UpU5GWD	CACGCCACGCTTCCTAATCG	PCR1 of Δgwd
3′-InDHFR	CAAGACGCAGACGCATACAA	PCR1 of Δusp and Δgwd
5′-InDHFR	CGCACGGACGAATCCAGATG	PCR2 of Δusp and Δgwd
3′-DnU3USP	TCAAAGACGACTGCATGC	PCR2 of Δusp
3′-DnU3GWD	AACCTTGCGCTTCAATCC	PCR2 of Δgwd
In-USP-Fw	CCTGGGATCTATTGCTGGA	PCR3 of Δusp
In-USP-Rv	GGCCTGCAGAGAAAAAGAA	PCR3 of Δusp
In-GWD-Fw	AACCTTGCGCTTCAATCC	PCR3 of Δgwd
In-GWD-Rv	TCCAGAAAGCCGATGACG	PCR3 of Δgwd
RT-tubulin-Fw	CACTGGTACACGGGTGAAGGT	β-tubulin-based qRT-PCR
RT-tubulin-Rw	ATTCTCCCTCTTCCTCTGCG	β-tubulin-based qRT-PCR
RT-USP-Fw	GTAGTTCTCGCTCCGTCCTG	USP-based qRT-PCR
RT-USP-Rw	CCGTCTTCAAAGGCCATCCT	USP-based qRT-PCR
RT-GWD-Fw	TCTGAAACGACGTTCCCCTG	GWD-based qRT-PCR
RT-GWD-Rw	TCACGGCGATGATAGTGTCG	GWD-based qRT-PCR

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Animal Diseases