The invention belongs to animal
biology technology and breeding field of pigs, and particularly relates to
cloning, activity analysis and
promoter region
mutation research of promoters of porcine
isocitrate dehydrogenase genes IDH3beta. The method comprises the following steps of: extracting genomes
DNA from
porcine blood, designing primers, obtaining
genome base sequences of upstream 5' flank 2447bp of porcine IDH3beta genes in two-time amplification mode by adopting
TAIL-PCR technology, and performing
sequence comparison and analysis and
genetic variation detection; and amplifying the upstream 5' flank region of the IDH3beta genes by adopting 5'-end deletion strategy, constructing
luciferase report
gene vectors by using the obtained
DNA sequences, measuring the activity of the promoters, and identifying the promoters of the IDH3beta genes. The
nucleotide sequences of the promoters of the cloned porcine
isocitrate dehydrogenase genes IDH3beta are expressed as sequence tables SEQ ID No: 1 and SEQ ID No: 4. The invention provides
gene resources for porcine genetic improvement application.