Primer used for clone DNA from
H. minnesotensis
|
5Sac-FP |
CGAGCTCCTGGTAAAATCTGTCCCG |
968 bp |
Amplified 5′ UTR and partial sequence of HIM-SKN7
|
This study |
5Xma-RP |
CCCCGGGCTGTCGGATAAAACTGGA |
This study |
3Asc-FP |
GGCGCGCCAATAAGGTGATGAAGCAA |
987 bp |
Amplified 3′ UTR and partial sequence of HIM-SKN7
|
This study |
3Pac-RP |
CTTAATTAAACCCAATGCGGCACAAGC |
This study |
SKN7OX-F |
TCTCAAGGTAGCGGCAACAAT |
1.8 kb |
Amplified HIM-SKN7 ORF for over-expression and to confirm replacement with hph cassete |
This study |
SKN7OX-R |
CCCGCACTGTCTCCATCGTGT |
This study |
Primer used for identification of null mutants |
5-HD-FP |
GCCGAGGACTTCACCACTA |
741 bp |
Amplified HIM-SKN7 gene (PCR 1) |
This study |
3-HD-RP |
GCAGCCCATTGACTTGAGA |
This study |
HD-F5N |
GGCAGTTCGTGGTTTAGTCG |
2.2 kb |
Amplified sequence of UTR and partial sequence of hph gene (PCR 2) |
This study |
Hyg-R3 |
CTTTGTAGAAACCATCGGCG |
This study |
Hyg-NF |
AAGTTCGACAGCGTCTCC |
937 bp |
Amplified the gene hph (PCR 3) |
This study |
Hyg-NR |
TTCCACTATCGGCGAGTA |
This study |
Primer used for identification of over-expression mutants |
PromSP3 |
AGGTAAGTGAACGACCCGGTC |
1891 bp |
Amplifying ORF and partial sequence of PtrpC (PCR4) |
This study |
SKN7OX-R |
CCCGCACTGTCTCCATCGTGT |
This study |
Hyg-NF |
AAGTTCGACAGCGTCTCC |
937 bp |
Amplified the gene hph (PCR 5) |
This study |
Hyg-NR |
TTCCACTATCGGCGAGTA |
This study |
Primer used for RT-PCR and RT-qPCT |
HRT-F |
ACCTAAGCCCATACAACG |
254 bp |
mRNA transcription of HIM-SKN7 gene |
This study |
HRT-R |
TAAATCATTCATCGGGTCAT |
HIM-08F |
GGAAACCCAGAAACGACCCT |
149 bp |
mRNA transcription of HIM_00008 gene |
This study |
HIM-08R |
AGTCGGGTTCGGCTTGTTGT |
HIM-29F |
TTCTGTATGGGCTGCGTCTC |
181 bp |
mRNA transcription of HIM_00329 gene |
This study |
HIM-29R |
TCTGGCTCATCAACAAACCC |
Actin-F |
CCCCATCTACGAGGGTTT |
554 bp |
Used as internal control |
This study |
Actin-R |
GGGAAGCGAGAATGGAAC |