Nucleic acid vaccine, and its application
A nucleic acid vaccine and vaccine technology, applied to nucleic acid vaccines and their application fields, can solve problems such as insignificant effects of piglets, and achieve the effects of improving disease resistance and growth rate of livestock, improving feed utilization efficiency, and reducing animal production costs.
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Embodiment 1
[0033] Embodiment 1, construction of somatostatin SS and porcine reproductive and respiratory syndrome virus envelope glycoprotein GP5 doublet carrier and SD protein and SS doublet carrier of pseudorabies virus
[0034] Blood was collected from the anterior vena cava of 3-week-old piglets, and porcine peripheral blood mononuclear cells were separated with leukocyte separation solution (Institute of Hematology, Chinese Academy of Medical Sciences) and the cell genome was extracted. Using the porcine peripheral blood mononuclear cell genome as a template, the following two pairs of primers were used: 5'-aag aat tct cca tgg aac tgg cca agt act tct t-3' and 5'-tttgcg aat tcg ggg tgc cat gg-3' and 5'-aaa cta gtc agg aac tgg cca ag-3' and 5'-caa cac tag tta ata aag cta aca gg-3'guided PCR amplification of porcine somatostatin gene, and the amplified products were named as SS180 (contains EcoR I sites at both ends), SS237 (contains Spe I sites at both ends). SS180 and SS237 were con...
Embodiment 2
[0039] Embodiment 2, animal experiments
[0040] In order to detect whether the construct of SS and GP5 or gD fusion gene plasmid DNA can produce virus-neutralizing antibodies, animal experiments were carried out using mice and pigs as immunized animals.
[0041] The specific steps of immunizing mice are as follows: select 30 female BALB / c mice aged 3 weeks, feed them for 1 week and confirm their health, and divide them into 6 groups randomly, 5 mice in each group. After grouping, there was no difference in average body weight between groups (P>0.05), and the 6 groups of mice were randomly divided into groups 1, 2, 3, 4, 5, and 6, and were immunized with pISG-SS / GP5, pISG-GP5 / SS, and pISG respectively -SS / gD, pISG-gD / SS, pISGRTK3 empty vector plasmid DNA and saline. Groups 1-5 were intramuscularly injected with the corresponding plasmid DNA at a dose of 100 μg per mouse, and Group 6 was injected with an equal volume of 0.9% normal saline. Vaccine once every two weeks, a tota...
Embodiment 3
[0047] Embodiment 3, detecting that the nucleic acid vaccine of the present invention improves the weight gain ability of pigs
[0048] In this example, pigs were purchased from the experimental pasture of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, and 20 healthy weaned piglets of about one month old were selected, and no anti-PRRSV and PRV antibodies were detected by serological testing. 5 pigs with the same age and body weight were selected as a repetition, and a total of 4 repetitions were set up. One head was randomly selected for each repetition and divided into 1 group, with 4 heads in each group, 5 groups in total. After grouping, there was no difference in the average body weight between the groups (P>0.05), so the five groups of pigs were randomly divided into groups 1, 2, 3, 4, and 5, and were immunized with pISG-SS / GP5, pISG-GP5 / SS, pISG- SS / gD, pISG-gD / SS, pISGRTK3 empty vector plasmid DNA. Each pig in groups 1-5 was injected ...
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