Virus recombinant vaccine A-NDV-LX/14 for newcastle disease and construction method thereof
A technology of A-NDV-LX and Newcastle disease, applied in the direction of virus/bacteriophage, antiviral agents, virus antigen components, etc., can solve the problem of genotype inconsistency between strains and epidemic strains, and achieve the effect of broad application prospects
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Embodiment 1
[0024] Example 1. Construction of full-length clone prLX of NDV / LX strain.
[0025] build mode like figure 1 shown.
[0026] Step 1: Obtaining NDV / LX strain
[0027] A batch of attenuated Newcastle disease virus strains were isolated from the intestinal tract of healthy poultry, and their biological characteristics: hemagglutination value, ICPI, EID50 were analyzed, and among these attenuated strains, vaccine candidates with high hemorrhagic agglutination value and strong replication ability were screened and carry out immunogenicity analysis to the candidate strains, immunize 2-week-old SPF chickens, measure anti-Newcastle disease virus titers in the serum after immunization and the immune level of the digestive tract and respiratory tract mucosa, and finally obtain good in the respiratory tract and digestive tract Replication, induction of high-titer specific antibodies and mucosal immune response to Newcastle disease attenuated strain NDV / LX (genotype I Newcastle disease...
Embodiment 2
[0047] Construction Step 1: Construction of recombinant plasmid pA-NDV-LX / I4
[0048] 1. Construct a plasmid containing the F gene and the HN gene of the isolate NDV / LX. The primer sequences for amplification are as follows:
[0049] QDr2F: 5'-GCTCGAGACTCGGTATTCATCACCACCTAT-3' (SEQ ID NO.19) Xho I
[0050] LXrg2R: 5'-CGTCTCGGTCTACCTCCACATCAATAGTGAC-3' (SEQ ID NO.20) BsmB I
[0051] QDr3F: 5'-TACTTGAAGACGTAGACCCCGAAGA-3' (SEQ ID NO.21) Bbs I
[0052] QDr3R: 5'-GTCGACACAGATCATTATTTAGTGTCATGG-3' (SEQ ID NO.22) Sal I
[0053] LX3F2: 5'-GTCGACIACTTATAGTTAGTTCGCCTG-3' (SEQ ID NO.23) Sal I
[0054] 5'-CGTCTCGTCGAGATATCAAGATTGCCTGTCAC A-3' (SEQ ID
[0055] LXrg3R: BsmB I
[0056] NO.24)
[0057] LXrg4F5-ACGTCGACTGAGCTTGGGAATGTCAACAACTC-3 (SEQ ID NO.25) Sal I
[0058] LXrg4R 5-CAGTTGACTCAACCGGCTAGACCTGGCTTC-3 (SEQ ID NO. 26) Spe I
[0059] Using the clone prLX containing the full-length genome of the NDV / LX strain as a template, use primers QDr2F and LXrg2R to amplify the fragm...
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