Caspase-8 in Labeo rohita is evolutionary conserved and is activated in Aeromonas hydrophila and Edwardsiella tarda infection and rhabdovirus vaccination

The Journal of Basic and Applied Zoology

Table 1 Primers, their sequence, and application in this study

Primer	Sequence (5′ → 3′)	Application
(a) Primers used for PCR amplification and cloning of rohu caspase-8
Casp8-FW^#	CCAGCTGAAGTTTCTGTGC	Cloning
Casp8-RV^#	CCGAGATTGTCTTCTGTCATGTC
Casp8-FW^#	AMTATCTGAAGACATGACAGAAGA
Casp8-RV^#	CCCS TAGAGGR AGTCTGCCTCCT
Casp8-FW*	GGGCACAGTGTTGGGTATTGAT
Casp8-RV*	GTGCTCTACTGTGGCCATTCCTA
GSP-RV-5′	CCTTGTTCCTGGGCCCTGTAGTCCTGTATCCT	5′ RACE
GS-NP-RV-5′	GAGGCGTCACGTTGTTGCAGATTTCTTTCCAC	5′ Nested RACE
GSP-FW-3′	TGACAAAGGTAAACCATGAAGTGAGCT	3′ RACE
GS-NP-FW-3′	CAGTGTTGGGTATTGATGCTAAAGAGGTTGA	3′ Nested RACE
UPM	Long: CTAATACGACTCACTATAGGGCAAGCAGT GGTATCAACGCAGAGT	RACE
UPM	Short: CTAATACGACTCACTATAGGGC	RACE
NUP	AAGCAGTGGTATCAACGCAGAGT	Nested RACE
(b) Primers used for rohu-caspase-8 gene expression analysis by quantitative real-time PCR (qRT-PCR) assay
Casp-8 FW	GGGCACAGTGTTGGGTATTGAT	Real-time PCR
Casp-8 RV	GTGCTCTACTGTGGCCATTCCTA
β-actin FW	AGACCACCTTCAACTCCATCATG
β-actin RV	TCCGATCCAGACAGAGTATTTACGC

Primers marked with “#” were designed from the conserved regions of caspase-8 cDNA sequence of Danio rerio (acc. no. AF273220.1), Ctenopharyngodon idella (acc. no. KP145003.1), and Cyprinus carpio (KC822471.1), and primers “*” have been designed from the transcriptome sequence of rohu caspase-8. “M” stands for A/C, “S” for G/C, “R” for A/G nucleotide