Enhanced and Secretory Expression of Human Granulocyte Colony Stimulating Factor by Bacillus subtilis SCK6

Table 1

Sequence of oligonucleotides used to amplify insert (IF/IR) and vector (VF/VR) DNAs during prolonged overlap extension (POE) PCR.

Primer	Sequence 5′-3′
VF	CCTTGCCCAGCCCTGATAGAAGCTTGGATCCGGAGTCGAACCATAAAAGC
VR	TGGCAGGGCCCAGGGGGGTCATATGAGCTGATGCCGAATACGTAAAGGTA
IF	TACCTTTACGTATTCGGCATCAGCTCATATGACACCTCTGGGCCCTGCCA
IR	GCTTTTATGGTTCGACTCCGGATCCAAGCTTCTATCAGGGCTGGGCAAGG

Primers were designed using online available software (http://www.xiaozhouzhang.com). AAGCTT and CATATG (underlined sequences) are the recognition sites for the HindIII and NdeI restriction endonucleases, respectively.