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Table 1 Primer sets used in the present study

From: Glucose- and glutamine-driven de novo nucleotide synthesis facilitates WSSV replication in shrimp

Gene

Primer

Primer sequence (5’-3’)

Function

G6PDH (PVHP204597.4)

G6PDH-qF

5’-CGCCACCATCAACAACGA-3’

Real-time PCR

G6PDH-qR

5’-AGGGCCTTTCCGCATCTG-3’

Real-time PCR

G6PDH-dsF

5’-AAGCTGTCGAACCACCTG-3’

Cloning

TKT

(PVHP71465.2)

TKT-qF

5’-CACTGCCTTCCAGCATGACA-3’

Real-time PCR

TKT-qR

5’-ATTCCAGCCAAAACCTTCGA-3’

Real-time PCR

TKT-dsR

5’-GAGACAAAAGCAACAGTACGGT-3’

Cloning

TKT-qF-T7

5’-TAATACGACTCACTATAGGGAGACACTGCCTTCCAGCATGACA-3’

dsRNA synthesis

TKT-dsR-T7

5’-TAATACGACTCACTATAGGGAGAGAGACAAAAGCAACAGTACGGT-3’

dsRNA synthesis

PRPS

(PVHP205382.2)

PRPS-qF

5’-TCTATCGCGAGCCCCAATT-3’

Real-time PCR

PRPS-qR

5’-GGCCACGGAGAGCATGTT-3’

Real-time PCR

PRPS-dsR

5’-AGACTCCTTCATGTGCTG-3’

Cloning

PRPS-qF-T7

5’-TAATACGACTCACTATAGGGAGATCTATCGCGAGCCCCAATT-3’

dsRNA synthesis

PRPS-dsR-T7

5’-TAATACGACTCACTATAGGGAGAAGACTCCTTCATGTGCTG-3’

dsRNA synthesis

ATase

(PVHP244493.1, PVHP244501.1)

Atase-qF

5’-TCCGTTGATGCTGATATTATTTCC-3’

Real-time PCR

Atase-qR

5’-ATCCCATAGCAGCCGGAGTA-3’

Real-time PCR

ADSS

(PVHP165389.1)

ADSS-qF

5’-CGGATGCGTCCCCTTGT-3’

Real-time PCR

ADSS-qR

5’-GTTGCTATGGAGACACTTGTGGAT-3’

Real-time PCR

IMPDH (PVHP227689.1)

IMPDH-qF

5’-AAGCACAGGAGGTCATGAAAGTC-3’

Real-time PCR

IMPDH-qR

5’-GGCAAACAGGGTCACGAATG-3’

Real-time PCR

IMPDH-dsR

5’-TCAACACCAGCATTGACAAG-3’

Cloning

IMPDH-qF-T7

5’-TAATACGACTCACTATAGGGAGAAAGCACAGGAGGTCATGAAAGTC-3’

dsRNA synthesis

IMPDH-dsR-T7

5’-TAATACGACTCACTATAGGGAGATCAACACCAGCATTGACAAG-3’

dsRNA synthesis

CAD

(PVHP274591.1)

CAD-qF

5’-CGGTTCAAGTTTTCGCGTATG-3’

Real-time PCR

CAD-qR-F2-dsR

5’-TGGGCTGACTTGAGGTTGGT-3’

Real-time PCR

CAD-F1-dsF

5’-CATTGGTCCTCCGTCAAT-3’

Cloning

CAD-F1-dsR

5’-CCGTGATTCTGAGACGTG-3’

Cloning

CAD-F2-dsF

5’-AGAAGCACAACATCATGCCT-3’

Cloning

CAD-F3-dsF

5’-AACATGTCCTGAGTGTTC-3’

Cloning

CAD-F3-dsR

5’-GCATACTGTGTGACATAG-3’

Cloning

CAD-F1-dsF-T7

5’-TAATACGACTCACTATAGGGAGACATTGGTCCTCCGTCAAT-3’

dsRNA synthesis

CAD-F1-dsR-T7

5’-TAATACGACTCACTATAGGGAGACCGTGATTCTGAGACGTG-3’

dsRNA synthesis

CAD-F2-dsF-T7

5’-TAATACGACTCACTATAGGGAGAAGAAGCACAACATCATGCCT-3’

dsRNA synthesis

CAD-F2-dsR-T7

5’-TAATACGACTCACTATAGGGAGATGGGCTGACTTGAGGTTGGT-3’

dsRNA synthesis

CAD-F3-dsF-T7

5’-TAATACGACTCACTATAGGGAGAAACATGTCCTGAGTGTTC-3’

dsRNA synthesis

CAD-F3-dsR-T7

5’-TAATACGACTCACTATAGGGAGAGCATACTGTGTGACATAG-3’

dsRNA synthesis

DHODH

(PVHP148100.1)

DHODH-qF

5’-GGCAGCAGGATTTGATAAGCAT-3’

Real-time PCR

DHODH-qR

5’-AAAGCTGAATCCCATACGAAACA-3’

Real-time PCR

DHODH-dsR

5’-AGAAACGGTTGTGTTGCTT-3’

Cloning

DHODH-qF-T7

5’-TAATACGACTCACTATAGGGAGAGGCAGCAGGATTTGATAAGCAT-3’

dsRNA synthesis

DHODH-dsR-T7

5’-TAATACGACTCACTATAGGGAGAAGAAACGGTTGTGTTGCTT-3’

dsRNA synthesis

UMPS

(PVHP164355.1)

UMPS-qF-dsF

5’-AACTCATGTGGGCCAAAC-3’

Real-time PCR

UMPS-qR

5’-GGCTACCCCGCAAACCAG-3’

Real-time PCR

UMPS-dsR

5’-TAACTCCGCAGAGGTGGT-3’

Cloning

UMPS-dsF-T7

5’-TAATACGACTCACTATAGGGAGAAACTCATGTGGGCCAAAC-3’

dsRNA synthesis

UMPS-dsR-T7

5’-TAATACGACTCACTATAGGGAGATAACTCCGCAGAGGTGGT-3’

dsRNA synthesis

CMPK

(PVHP117215.1)

CMPK-qF

5’-CCTTGTTAGAACGTGCCATGAA-3’

Real-time PCR

CMPK-qR

5’-CCGAGGAAATCCATCAATGAG-3’

Real-time PCR

CMPK-dsF

5’-CCAAGCTTATGTTTTTGAATCGAATT-3’

Cloning

CMPK-dsR

5’-CCGGATCCACTGAACAGCTTTTCAAC-3’

Cloning

CMPK-dsF-T7

5’-TAATACGACTCACTATAGGGAGAATGTTTTTGAATCGAATT-3’

dsRNA synthesis

CMPK-dsR-T7

5’-TAATACGACTCACTATAGGGAGAACTGAACAGCTTTTCAAC-3’

dsRNA synthesis

Luciferase

Luc-F

5’-CTGAATACAAATCACAGAATC-3’

Cloning

Luc-R

5’-GCGAGAATCTGACGCAGGCAGT-3’

Cloning

Luc-F-T7

5’-TAATACGACTCACTATAGGGAGACTGAATACAAATCACAGAATC-3’

dsRNA synthesis

Luc-R-T7

5’-TAATACGACTCACTATAGGGAGAGCGAGAATCTGACGCAGGCAGT-3’

dsRNA synthesis

EF-1α

EF-1α-qF

5’-ACGTGTCCGTGAAGGATCTGAA-3’

Real-time PCR

EF-1α-qR

5’-TCCTTGGCAGGGTCGTTCTT-3’

Real-time PCR

WSSV IE1

IE1-qF

5’-CCAGGCCCAGTGTCATACG-3’

Real-time PCR

IE1-qR

5’-AGAAATCTCATCACATGTCAAATCAGA-3’

Real-time PCR

WSSV VP28

VP28-qF

5’-AGTTGGCACCTTTGTGTGTGGTA-3’

Real-time PCR

VP28-qR

5’-TTTCCACCGGCGGTAGCT-3’

Real-time PCR

Others

Anchor-dTV

5’-GACCACGCGTATCGATGTCGACTTTTTTTTTTTTTTTTV-3’

Reverse transcription

  1. The primer set for the genes involved in de novo nucleotide synthesis was designed using an in-house L. vannamei transcriptomic database. Accession numbers are given below in parentheses.
  2. The added T7 promoter sequence is in italics.