TY - JOUR
T1 - Porcine tumor necrosis factor alpha
T2 - cloning with the polymerase chain reaction and determination of the nucleotide sequence
AU - Pauli, Urs
AU - Beutler, Bruce
AU - Peterhans, Ernst
PY - 1989/9/1
Y1 - 1989/9/1
N2 - We have cloned the gene for the porcine tumor necrosis factor α (TNF-α.) utilizing the polymerase chain reaction (PCR). Total RNA from stimulated monocytes was used to generate TNF-α-specific single-stranded cDNA, with reverse transcriptase and a conserved consensus primer, starting at the TNF-α. translation stop codon. After adding a second conserved consensus primer to the propeptide region, we amplified the cDNA between the two primers. The isolated fragment was cloned and sequenced. Comparison of the nucleotide sequence indicated an 85 % sequence similarity to the human TNF-α gene. Eighteen amino acids of the deduced mature peptide sequence of porcine TNF-α were different from those in the sequence of man. The technique used in this work allows rapid cloning of specific genes from total RNA, and, additionally, screens for full-length transcripts during the amplification procedure.
AB - We have cloned the gene for the porcine tumor necrosis factor α (TNF-α.) utilizing the polymerase chain reaction (PCR). Total RNA from stimulated monocytes was used to generate TNF-α-specific single-stranded cDNA, with reverse transcriptase and a conserved consensus primer, starting at the TNF-α. translation stop codon. After adding a second conserved consensus primer to the propeptide region, we amplified the cDNA between the two primers. The isolated fragment was cloned and sequenced. Comparison of the nucleotide sequence indicated an 85 % sequence similarity to the human TNF-α gene. Eighteen amino acids of the deduced mature peptide sequence of porcine TNF-α were different from those in the sequence of man. The technique used in this work allows rapid cloning of specific genes from total RNA, and, additionally, screens for full-length transcripts during the amplification procedure.
KW - Recombinant DNA
KW - amino acid sequence homology
KW - amplification
KW - cachectin
KW - complementary DNA
KW - evolution
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U2 - 10.1016/0378-1119(89)90350-8
DO - 10.1016/0378-1119(89)90350-8
M3 - Article
C2 - 2478420
AN - SCOPUS:0024464078
SN - 0378-1119
VL - 81
SP - 185
EP - 191
JO - Gene
JF - Gene
IS - 1
ER -