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Table 2 Primers used for SDM and cloning of the MHV spike and IBV nucleocapsid gene

From: A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination

Primer

Sequence (5′ ➔ 3′)

Function

XhoI-t-MHV-S.F01

CTCGAG TTATATTGGTGATTTTAGATGTATCCAG

Cloning MHV S

MHV-S-StyI-XhoI.R01

CTCGAGCCAAGGCCATTTCACATACATTTC

Cloning MHV S

p-MHV-S-SDM414.F02

AGCTTGTGAACTCAAACGGTG

SDM EcoRI

p-MHV-S-SDM414.R02

GGATACATCTAAAATCACCAATATAAC

SDM EcoRI

p-MHV-S-SDM2823.F03

TTACTATAAGTTCGAGACTGCC

SDM XhoI

p-MHV-S-SDM2823.R03

CACCCTGCATTAATGCAC

SDM XhoI

IBV-H52_N_ATG_FW

ACCATGGCGAGCGGTAAGA

N-transcript

IBV-M41-#2-IR-RV

TTTTTTTTTTTTTTTTTTTGCTCTAACTCTATACTAG

N-transcript

  1. XhoI restriction enzyme sites in sequences of primers XhoI-t-MHV-S.F01 and MHV-S-StyI-XhoI.R01 are in italics; an additional thymidine residue to keep the MHV spike gene ectodomain sequence in frame with the IBV spike gene signal sequence is in bold. SDM = site directed mutagenesis
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Virology Journal

ISSN: 1743-422X