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In this study, has been constructed pcDNAKCpG-1, 2,3,4, and every hole 1.25μg, 2.5μg, 5.0μg, 10.0μg doses stimulate lymphocyte separation medium with an SPF pig peripheral blood mononuclear cells (PBMC ) , carried out in vitro lymphocyte transformation test , transformed by MTT colorimetric assay results, to compare the different plasmids and their concentration on the stimulatory effects of porcine peripheral blood lymphocytes . The results showed that: four kinds of plasmid on pig peripheral blood mononuclear cells have different levels of stimulation effects, which 10.0μg pcDNAKCpG-4 doses of swine lymphocyte stimulation index reached 6.3; pcDNAKCpG-2 at lower doses it has higher stimulation index , the pig has the best immunostimulatory activity . With the pig has the best immune stimulating activity of recombinant plasmid pcDNKCpG-2, according to 500μg / head muscles dose inoculated pigs , respectively, after injection 4h, 4d, 8d, 15d, 30d slaughter and collecting , extracting major tissues and organs DNA, then the recombinant plasmid sequences by PCR primers to identify the recombinant plasmid in pigs and residence time distribution . The results showed that: after the injection 4h, the site of injection in pig muscle detected recombinant plasmid pcDNAKCpG-2; after injection 4d, in a pig 's blood and injection site muscle detect recombinant plasmid pcDNAKCpG-2; 8d after injection that any organization can not detect the presence of recombinant plasmids . Based on these studies , further positive PCR test swine major organizations gDNA and 30d after injection of porcine major organizations gDNA, with digoxin labeled CpG sequences as probes for Southern blotting test detects whether CpG sequences pig gDNA integration occurs . The results showed that : CpG sequences and porcine gDNA integration did not occur . In short, the recombinant plasmid pcDNAKCpG-2 on porcine immune stimulation has the best results, as an immune adjuvant in pigs residence time of less than 8d, and does not occur with the pig genome integration .
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