Table 1.

Primers Utilized to Synthesize Chicken GM-CSF

Primers for the first PCR reaction:

Forward primer #1 (HindIII-Kozak-ORF-nucleotides #1–80): 5′-GCAT AAGCTT CCACG ATGCTGGCCC AGCTCACTAT TCTGCTTGCC CTCGGGGTGC TCTGCAGCCC TGCGCCCACC ACAACATACT CCTGCTGCTA-3′

Reverse primer #2 (nucleotides # 155 to 156): 5′-ACCGA GGACAGACCT GCTGTGGCCG CTGTGCTCTC CAAGTGACTC GTTATTTCTT CCAGGATGGT GTACACTTTG TAGCAGCAGG AGTATGTTGT GGTGG-3′

Forward primer #3 (nucleotides # 131–230): 5′- CGGCCACAGC AGGTCTGTCC TCGGTACCCA TGGACATCAG GGATAAAACC TGTCTGCGTA ACAACCTGAA AACATTCATA GAGTCCTTGA AAACAAATGG-3′

Reverse primer #4 (nucleotides # 305–206): 5′-GTTAT GTTCGAGAAG AGGCGTTCAC ACTCGTGAAC TCTGTTCAGC TGAAAGACGA TTCCGCTTTC TTCCTCTGTC CCATTTGTTT TCAAGGACTC TATGA-3′

Forward primer #5 (nucleotides # 281–380): 5′-GTGAACGCCT CTTCTCGAAC ATAACTCCCA CCCCGCAGGT TCCTGATAAG GAATGTAGAA CTGCACAAGT ATCGAGGGAA AAATTCAAAG AGGCATTAAA-3′

Reverse primer #6 (nucleotides # 435–356): 5′-GCAT GAATTC TTAGA TGCAGTCTTT CTCCTCTGGG AGCACATCAG AGAGGTAAAT AAAGAAAGTT TTTAATGCCT CTTTGAATTT TTCCC-3′

Primers for the second PCR reactions to make soluble and membrane-bound GM-CSF:

Primer #7 (used for soluble and membrane-bound GM-CSF) (forward primer with HindIII restriction site): 5′-GCAT AAGCTT CCACG ATGCTGGCCC AGCTC-3′

Primer #8 (used for soluble GM-CSF) (reverse primer with EcoR1 restriction site): 5′-GCAT GAATTC TTAGA TGCAGTCTTT CTCCT-3′

Primer #9 (used for membrane-bound GM-CSF) (reverse primer with BamH1 restriction site): 5′-GCATAAGCTTCCACGATGCTGGCCCAGCTC-3′