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Table 3 Forward and reverse primers used to amplify specific kinase genes. Note also preferred vector for each construct

From: Novel kinase platform for the validation of the anti-tubercular activities of Pelargonium sidoides (Geraniaceae)

Kinase

Gene and Rv identifier

Forward and Reverse primers (5′ → 3′)

Restri-ction Enzyme Recogni-tion site (underli-ned)

Selected pET vector

NDK

ndkA Rv2445c

ndkA-Fwd

 

pET-16b

GGCATATGACCGAACGGACTCTGGTACTG

NdeI

 

ndkA-Rev

GTGGATCCTTAGGCGCCGGGAAACCAG

BamHI

 

HSK

thrB Rv1296

thrB-Fwd

 

pET-20a

GGCATATGGTGACTCAAGCATTG

NdeI

 

thrB-Rev

GTCTCGAGACCGGGAACTCTTACTG

XhoI

 

AK

ackA

Rv0409

ackA-Fwd

 

pET-16b

GGCATATGGAGTAGCACCGTGCTGGTGATCAA

NdeI

 

ackA-Rev

GTGGATCCTTACGCTCGGCGTCCGCCCAG

BamHI

 

GK

glpK

Rv3696c

glpK-Fwd

 

pET-16b

(GGCATATGTCCGACGCCATCCTAG

NdeI

 

glpK-Rev

CATGTCGACTTAGGACACGTCAACCCAATCC

SalI

 

ThiL

thil

Rv2977c

thil-Fwd

 

pET-28a

GGTACATATGACCACTAAAGATCACTC

NdeI

 

thil-Rev

GATCTCGAGTTACCCTAGCGAACCTTG

XhoI

 

RBKS

rbks

Rv2436

rbks-Fwd

 

pET-28a

GTACATATGGCAAACGCCAGTGAG

NdeI

 

rbks-Rev

GATCTCGAGTTATGAACCGTTGTG

XhoI

 

AsKa

ask

Rv3709c

ask alpha-Fwd

 

pCDF-

GATTACATATGGCGCTCGTCGTGCAG

NdeI

Duet-1

ask alpha-Rev

 

(alpha)

GATGTCGACTTACCGTCCCGTCCCCG-3’

SalI

 

ask beta-Fwd

 

pET-26a

GATCATATGGAAGACCCCATCCTGACCG

NdeI

(beta)

ask beta-Rev

  

GCCCCTGCCCTGCCCAGCTGTATG

SalI

 

SK

aroK

Rv2539c

Primers were not designed as the aroK plasmid was received from the laboratory of Chris Abell, University of Cambridge

 

pET-15b

  1. a AsK consists of 2 hetero-monomers (alpha and beta), to be co-expressed