| Porcine reproductive and respiratory syndrome (PRRS) was a disease which was first described to occur in the United States in 1987 and in Europe in 1991. Its current economic impact estimated at million annually. Different isolates of PRRSV exist nucleotide sequence variation and bring to difficuties on PRRS accurate detection and effective control. This study aimed to construct and express the NSP2 of hypervarible and American strain, in addition,establish the ELISA method,for the development of effective prevention and control of PRRSV immune to provide reasonable basis.According to published GenBank sequences of VR-2332 strain, NSP2 gene for this study was designed two pairs of primers; using RT-PCR method successfully amplified the separation of TA-12 strain NSP2 gene. DNAStar software analysis NSP2 protein epitope were divided into four, N-conservative region into two fragments, the middle hypevariable region to a fragment, C-end fragment as a conservative region, designed four pairs of restriction sites with primers, respectively, the recombinant plasmid was constructed, Western blot identification results showed that recombinant protein express in the host bacteria. At the same time, the use of TA-12NSP2 amplified hypervariable region of the RT-PCR primers by the Inter-American strain VR-2332 NSP2 of the corresponding gene fragment, successfully inserted into the plasmid of the correct fragment in pET-28a. This experiment a further option to the TA-12 conserved region and the middle hypevariable region purified protein, and the establishment of ELISA method of detection of the 400 collected in this experiment serum of pigs. ELISA results showed that PRRSV NSP2 region gene mutant among the TA-12 and the Inter-American standard strains with different epitope immunogenicity that can be used to distinguish PRRSV protein NSP2 mutant strain with the standard for precision PRRSV immune to provide a basis for monitoring. |
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