Development an effective system to expression recombinant protein in E. coli via comparison and optimization of signal peptides: Expression of Pseudomonas fluorescens BJ-10 thermostable lipase as case study

Table 1 Oligonucleotides used in the experiments

Oligonucleotides	Sequence (5′–3′)
PFL-22b (upstream)^a	CCGCATATGGGTGTCTACGACTACAAAAACC
PFL-22b (downstream)^a	CCGCTCGAGGGTAATCACAAACGCCTCCG
PFL-SP-22b (upstream)^b	CCGGAATCCATGGGTGTCTACGACTA
PFL-SP-22b (downstream)^b	CCGCTCGAGGGTAATCACAAACGCCT
pET-PFL (upstream)^c	GGGGAATTGTGAGCGGATAAC
pET-PFL (downstream)^c	TGGCAGCAGCCAACTCAG
16s-reference (upstream)^d	AATCATCATGCCCCTTATGACC
16s-reference (downstream)^d	GTTGCAGCCTACAATCCGAAC
PFL-target (upstream)^e	GGTGGAAGTCCTGGGCAAAT
PFL-target (downstream)^e	CGCCGATGGAATCAACAA

^aPrimers PFL-22b were used to amplify the full-length sequence of lipase gene lipBJ10, with NdeI and XhoI restriction sites (underlined), respectively
^bFor fusion with six different signal peptides, primers PFL-SP-22b were used to amplify the full-length sequence of lipase gene lipBJ10, with EcoRI and XhoI restriction sites (underlined), respectively
^cPrimers pET-PFL were used to verify the sequence of recombinant plasmid
^dPrimers 16s-reference were used to amplify E. coli 16s sequence
^ePrimers PFL-target were designed for lipBJ10

ISSN: 1475-2859