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Table 1 Oligonucleotides used in this study

From: Modification of genetic regulation of a heterologous chitosanase gene in Streptomyces lividans TK24 leads to chitosanase production in the absence of chitosan

Aim of primers

Name

Sequence (5'→3')

For csnN106 coding region cloning*

FwcsnN106

CCGGAGACCCGCATGC CCCGGAC

 

RvcsnN106

CGGTGCGCCAAGCTT GCGTTCGG

For Pr-WT cloning*

FwPr-WT

GTCTGCGCGGATCC TGACGGCCC

 

RvPr-WT

GTCCGGGGCATGC GGGTCTCCGG

PCR-directed mutagenesis for Pr-Pa cloning**

SEQ.1

ACAACTTCGTCGCGCACATCCA

 

Rw1Pr-Pa

ATGAGGAGAGTTCGGACAGTTTC

 

Fw2Pr-Pa

GAAACTG TCCG AACTCTCCTCAT

 

RvcsnN106

TGAGGTCGAAGTTCTTGGCGTT

Verification of pHM8a derivatives integration into hosts

Fwgenom

CCTGAGAGGCCGGTGAGGAG

 

RvcsnN106

TGAGGTCGAAGTTCTTGGCGTT

Presence verification of pFDES derivatives into hosts

SEQ.1

ACAACTTCGTCGCGCACATCCA

 

T7 promoter

TTAATACGACTCACTATAGGG

For Primer extension

PE-csnN106

TGGGGTGCTTGAGACGCAT

  1. *Bold nucleotides correspond to restriction site
  2. **Bold nucleotide correspond to mutated nucleotide