TABLE 1

Primers used for PCR amplification and BNBD3-encoding complementary synthetic oligonucleotide pairs

NamePrimer or oligonucleotide sequence
BNBD3-15′-GATCCATATGCAAGGAGTAAGAAATCATGTAACCTGCCGTATAAATAGAGGCT-3′a
BNBD3-1c5′-CAGAAGCCTCTATTTATACGGCAGGTTACATGATTTCTTACTCCTTGCATATG-3′a
BNBD3-25′-TCTGTGTGCCGATCAGGTGCCCTGGACGCACGAGACAGATTGGCACCTGTTTC-3′b
BNBD3-2c5′-GCCCGAAACAGGTGCCAATCTGTCTCGTGCGTCCAGGGCACCTGATCGGCACA-3′b
BNBD3nL-35′-GGGCCCCGAATAAAATGCTGCAGGTCGTGGTAGA-3′c
BNBD3nL-3c5′-AGCTTCTACCACGACCTGCAGCATTTTATTCGGG-3′c
BNBD3L-35′-GGGCCCCGAATAAAATGCTGCAGGTCGTGGAACGACGCCCAGGCCCCCAAGAGCA-3′d
BNBD3L-3c5′-AGCTTGCTCTTGGGGGCCTGGGCGTCGTTCCACGACCTGCAGCATTTTATTCGGG-3′d
  • a First piece of BNBD3; a 5′ BamHI restriction site is underlined, and the start codon is in bold.

  • b Second piece of BNBD3; oligonucleotides were 5′ phosphorylated, and sticky ends were made by a 4-nucleotide overhang (italic).

  • c Third piece of BNBD3 (pMASIA-BNBD3); a 3′ HindIII restriction site is underlined, and the TAG stop codon is in bold.

  • d Third piece of BNBD3 (pMASIA-BNBD3-tgD); the bovine codon optimized linker is in italic, and a 3′ HindIII restriction site is underlined.