廖仁寶 張秀鑾 吳明哲
SEQUENCING OF THE PROMOTER REGION OF
GENE IN BERKSHIRE PIGS
R. B. Liaw, H. L. Chang and M. C. Wu
Taiwan Livestock Research Institute
There is a variable number of GT dinucleotide repeats in the promoter region of osteopontin (OPN) gene in pigs. The fragments of the promoter region of osteopontin gene from four Berkshire pigs were amplified with PCR. Then the amplicons were ligated to the cloning vector and transformed to Escherichia coli. There were 4, 4, 3, and 5 white colonies, respectively, selected from the four pigs. Recombinant plasmids were extracted from transformants and inserts were sequenced by an automated DNA sequencer. The insert sequence was 5’CCAATCCTATTCACGAAAAAGCTAGTTAATGACATTGTACATAAGTAATGTTTTAACTGCAGAGGGC(GT)nTGTAACCACAAAACCAGAGGGGGAAGTGTGGGAGCAAGTGGGTTG3’ (the forward and reverse primers were in bold and italic). All insert sequences of recombinant plasmids were the same except of the number of GT dinucleotide repeats. Number of GT repeats of the inserts from B9510806 pig was 14 only. GT repeats of the inserts from B9560505 pig were 13, 14 and 24. GT repeats of the inserts from B100303 pig were 24 and 25. GT repeats of the inserts from B100501 pig were 22, 23, 24 and 25. Results from this study indicated that there were six alleles of the 13, 14, 22, 23, 24 or 25 repeats of GT in the promoter region of osteopontin gene based upon four pigs of Berkshire breed.
Key Words: Berkshire, Osteopontin, Sequencing.