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Image Search Results
Journal: Frontiers in Veterinary Science
Article Title: Rapid Development of an Effective Newcastle Disease Virus Vaccine Candidate by Attenuation of a Genotype VII Velogenic Isolate Using a Simple Infectious Cloning System
doi: 10.3389/fvets.2020.00648
Figure Lengend Snippet: Genome organization of NDH3 and the schematic strategy for construction of the full-length pBR322-NHD3 and mutant pBR322-NHD3-mF constructs. (A) Ten overlapping PCR fragments covering the whole DHN3 genome are indicated by N, MIMI, PD, PD1, PD2, PD3, L1, L2, L3, and L4, respectively. The three subgenome-sized plasmids, pBR322-PNP, pBR322-PDP, and pBR322-LPD3, were constructed based on the indicated restriction sites and used for the construction of pBR322-DHN3. (B) The pBR322-Base plasmid was constructed by inserting a synthetic DNA fragment into pBR322 between Not I and Nhe I site. The DNA fragment is composed of the T7 terminator, HDV ribozyme, HC2 (5′ end from 1-141nt of DHN3), HC1 (3′ end from 15192-15159nt of DHN3) and the T7 promoter sequence. The PNP, PDP and LPD3 fragments excised from the three subgenome-sized plasmids were ligated, and then recombined with pBR322-Base to produce the full-length pBR322-DHN3. (C) pBR322-DHN3-mF was constructed by recombination of three fragments. The large fragment prepared from SmaI- digestion pBR322-DHN3, and the two smaller fragments containing the mutated sequence marked in red were generated by PCR amplification with two pairs of specially designed primers .
Article Snippet: Construction of pBR322-Base: a DNA fragment containing the T7 promoter, T7 terminator, HDV (Hepatitis delta virus) Ribozyme,
Techniques: Mutagenesis, Construct, Plasmid Preparation, Sequencing, Generated, Amplification