Toxicity of DON on GPx1-Overexpressed or Knockdown Porcine Splenic Lymphocytes In Vitro and Protective Effects of Sodium Selenite

Figure 1

Establishment of GPx1 gene-overexpressing model of pig splenic lymphocytes. (a) shows the result of PCR amplification of the GPx1 gene of porcine splenic lymphocytes; it shows the PCR products, showing that the length of the single target gene was 621 bp. The PCR-amplified GPx1 gene was inserted into PEGFP-N1, a eukaryotic vector with kanamycin resistance, to obtain the recombinant plasmid pEGFP-N1-GPx1. Positive colonies were screened on kanamycin-containing plates. To avoid false positives, the selected colonies were screened by PCR, as shown in (b). The positive colonies were then verified by XhoI and SacII double-enzyme digestion to obtain bands of 630 bp and 4700 bp, as shown in (c). The X-tremeGENE HP DNA transfection reagent was used to transfect the purified plasmid into pig spleen lymphocytes over 48 h, and the efficiency of transfection was detected by fluorescence inverted microscopy, as shown in (d and e). (f) is the result of protein immunoblotting for GPx1. In (f), 1 is the untreated control group, 2 is the pEGFP-N1-transfected empty vector group, and 3 is the pEGFP-N1-GPx1-transfected overexpressed group.

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