HA virus
1.Objective To detect the virulence/attenuation level and nucleotide sequences of HAV(H2 strain) cultured in human fetal lung diploid cell KMB17.Methods Live attenuated HA vaccine virus H2M20K7(K7) was proliferated in KMB17 cells and passaged at 35 and 37℃, then the immune responses of different passages in common marmosets were observed, and the nucleotide sequences of them were analyzed.
2.The paper is divided into three parts:The first part is the identification of the AEV-HD strain used in this test: the general identification were conducted, such as physical and chemic characters, hemagglutination trial (HA) and the virus conformation. The results showd that the characters of AEV-HD were stable, its rivulence wasn’t influenced by Trypsin, Aether, Chloroform and temperature (57 ℃1 h), could not agglutinate gallinaceous erythrocyte.
3.The hemagglutinin protein(HA) gene of avian influenza virus was amplified from pVAX1-HA by PCR and sub-cloned into pcDNA3.1(+) vector. HA-CpG gene was also cloned into the pcDNA3.1(+).
6.The immunizing effect produced in mice by influenza virus surfaceanti-gen HA subunit with muramyl dipeptide ( MDP) adjuvant is identical to that of HA subunit with A1 ( OH )3 and Freund adjuvant.
7.A pseudorabies virus (PRV) transfer vector(pLTK-HA)containing HA gene of H3N2 SIV used tocotransfect with genomic DNA of PRV Bartha-K61 into Vero cells. After many cycles of blue plaguepurification and PCR identification,we got a PRV recombinant containing hemagglutinin (HA) gene ofH3N2 SIV and designated as rPRV -HA.
8.We used the same method as above-mentioned and obtained the recombinant fowlpox virus expressing antigen part of HA gene, rFPV-12LS-HAA.
9.RESULTS The HA titer of CPV XN 1 was 1∶1024~1∶4096 or TCID 50 10 -7.0 . CPV XN 1 had a typical CPV morphological character underelectron microscope, antibody molecules could be observed to absorb on the surface of virus particles.
10.In this study, the influenza virus A/Equine/Jingfang/74-1(HTNT)was propagated in SPF egg embryo, and RNA was extracted with trizol LS reagent, then HA gene was amplified, cloned and sequenced.
