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Table 1 Oligonucleotides employed to obtain the coding DNA sequence of hG-CSF*.

From: Human granulocyte colony stimulating factor (hG-CSF): cloning, overexpression, purification and characterization

P1-5' AA CATATG ACCCCCCTGGGCCCTGCCAGCTCCCTGCCCCAGAGCTTCCTGCTC AAGTG 3'

P2-5' GCGCTGCGCCATCGCCCTGGATCTTCCTCACTTGCTCTAAGCACTTGAGCA 3'

P3-5' GGCGCAGCGCTCCAGGAGAAGCTGTGTGCCACCTACAAGCT 3'

P4-5' CCAGAGAGTGTCCGAGCAGCACCAGCTCCTCGGGGTGGCACAGCTTGTAGG 3'

P5-5' CACTCTCTGGGCATCCCCTGGGCTCCCCTGAGCAGCTGCCCCAGCCAGGCCCTG CAG 3'

P6-5' GGTAGAGGAAAAGGCCGCTATGGAGTTGGCTCAAGCAGCCTGCCAGCTGCA GGGCC 3'

P7-5' TTCCTCTACCAGGGGCTCCTGCAGGCCCTGGAAGGGATCTCCCCCGAGTTGGGT C 3'

P8-5' TGGTGGCAAAGTCGGCGACGTCCAGCTGCAGTGTGTCCAAGGTGGGACCCAACT C 3'

P9'-5' TTTGCCACCACCATCTGGCAGCAGATGGAAGAACTGGGAATGGCCCCT GCC CT 3'

P10-5' AAAGCAGAGGCGAAGGCCGGCATGGCACCCTGGGTGGGCTGCAGGGCAGGGG 3'

P11-5' CCTCTGCTTTCCAGCGCCGGGCAGGAGGGGTCCTGGTTGCCTCCCATCTGC AGAGCTTCC 3'

P12-5'AA GGATCC TCAGGGCTGGGCAAGGTGGCGTAGAACGCGGTACGACACCTCCAGGAAGCTCTG 3'

  1. * Primer sequences for construction of hG-CSF. The G-CSF nucleotide sequence (Accession number NM_000759) of 522 bp, was divided into 12 DNA sequences of approximately 50 bp each, containing 10-bp overlapping regions at their 5'- and 3'-ends for adjacent oligonucleotides. The restriction sites, Nde I and Bam HI, are underlined in primers P1 and P12, respectively.
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Microbial Cell Factories

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