Table 1 Strains, plasmids and oligonucleotides used in this study
Strains | Description | Source |
|---|---|---|
E. coli Trans T1 | lacх74 recA1 deoR F – mcrA Δ (mrr-hsdRMS-mcrBC) ϕ80 lacZΔM15Δ araD139Δ (ara-leu)7697 galU galK | Transgen |
T1-ΔepiA | E .coli cell carrying disruption cassette ΔepiA | This study |
Agrobacetrium tumefaciens (AGL-1) | Carrying pCAMBIA3301 plasmid, KanR | [42] |
AGL-1-ΔepiA | AGL-1 cell carrying ΔepiA | This study |
Epicoccum sp. CPCC 400996 | Wild type strain | |
Δ epiA -5 | epiA deletant of CPCC 400996 | This study |
Δ epiA -11 | epiA deletant of CPCC 400996 | This study |
Plasmids | Description | Source |
|---|---|---|
pAg1-H3 | Vector for targeted gene replacement containing HYG gene as selection marker | [59] |
H3-epiA-Dn | 3′ prime arms of epiA gene cloned into pAg1-H3 | This study |
H3-ΔepiA | 5′ & 3′ prime arms of epiA gene cloned into pAg1-H3 | This study |
Oligonucleotides | Sequence (5′–3′) | Purpose |
|---|---|---|
Z01 (epiA-Up-F) | CGACTAGTTTCATGTCAGCATGTAGC | 5′ prime arm amplication |
Z02 (epiA-Up-R) | CATTAATTAATGAGTCACGAACCTATT | |
Z03 (epiA-Dn-F) | ATCTCGAGTCAAACGCCAAAATGAAC | 3′ prime arm amplication |
Z04 (epiA-Dn-R) | ATGGGCCCTGAGCCCACCCGTGCCGG | |
Z05 (epiA_In-F) | ATTGATGAAGGAGAAGAGGGAG | In deletion region characterization |
Z06 (epiA_In-R) | GGAAACCAGGTTCGCAGT | |
Z07 (cha-epiA-UF) | GGGGCTACAATCTTTACG | epiA 5′ prime characterization |
Z08 (cha-epiA-UR) | GATACCAGACAACCACCTT | |
Z09 (cha-epiA-DF) | GTAAGCGGCACCAATCTC | epiA 3′ prime characterization |
Z10 (cha-epiA-DR) | AGCAACATCCCATCACAA | |
Z11 (cha-hph-F) | GGGCAAAGGAATAGAGTAGATG | Hyg + gene characterization |
Z12 (cha-hph-R) | TGCAATAGGTCAGGCTCT |