用本室分离鉴定的猪伪狂犬病毒鄂A 强毒株接种仓鼠肾细胞（BHK221），制备病毒抗原液，毒价不低于10-6 TCID50/011ml。经一定浓度的甲醛溶液灭活后与油相佐剂乳化研制成油乳剂灭活油疫苗4批。本研究对该制品的安全性、免疫性进行了测定。对18g左右小白鼠接种013ml，初生仔猪、断奶仔猪及妊娠母猪加倍剂量注射，均未出现不良反应，安全性良好。对母猪的繁殖性能不产生影响。后备母猪及妊娠母猪血清中和抗体指数于免疫后21d 达到316 以上，间隔35d强免疫一次后，中和抗体指数可达1000以上。断奶仔猪及初生仔猪免疫后对强毒的攻击，保护率分别为100%及90.62%。

以从湖北某猪场分离鉴定的鄂A株为亲本，提取其基因组DNA，克隆含gG基因的SphⅠ/KpnⅠ片段，然后将LacZ基因融合到gG启动子下游，得到重组质粒pUSKZ，将重组质粒与鄂A株基因组共转染PK215细胞，待细胞完全病变后，在X2gal存在下，作蓝斑筛选纯化。经斑点杂交和PCR扩增证实得到的是基因型为gG-/LacZ+的重组伪狂犬病病毒。

本研究从含有伪狂犬病病毒（Pseudorabies virus,PRV）Ea株gD基因BamHI 616Kb片段的重组质粒pUCB7中分别亚克隆gD基因的上、下游片段，并对上游片段进一步改造，去掉gD基因上游的非编码序列，构建了含完整gD基因编码区的重组质粒pBRgDI，并利用基因内部的限制性内切酶位点，用内切酶KpnI和SalI酶切分析，证实了gD基因的可靠性和完整性。同时构建了测序质粒pSKgDSB和pSKgDS，并用双脱氧终止法进行测序。将测序结果与国外的Rice毒株进行比较，发现Ea株gD基因核苷酸序列有多处点突变和一处插入突变，在编码氨基酸残基水平上也有差异。

Glycoprotein gene gB, gC and gD of pseudorabies virus (PrV) strain Ea, which was isolated locally in Wuhan, were cloned from the viral genome DNA and expressed in vitro controlled by the major immediately-early promotor/enhancer of HCMV. In the presented paper, Balb/c mice, rabbits and piglets were vaccinated intramuscularly two times at 2-week interval wth those eukaryotic expression plasmid pcDB, pcDC and pcDD, respectively. The animals injected with pcDB, pcDC, pcDD or mix DNA developed anti-PrV antibodies. Neutralizing antibody titers obtained 2-5 log2, 2 weeks after the second vaccination. Cellular immune responses were also detected by lymphoproliferation assay and cytotoxic T lymphocyte (CTL) activity assay in all groups vaccinated with DNA. Immune responses elicited by DNA vaccines provided protections with different degrees against lethal dose PrV challenge. In mice, protections induced by pcDC, pcDD or mix DNA were 100%, similar to that by inactivated vaccine. Protections were more than 50% induced by pcDC, pcDD or mix DNA in rabbits. Protections induced by pcDB were the lowest among DNA immunization in mice or rabbits. However, pcDB could elicit the higher cellular responses in rabbits or piglets. In piglets, body temperatures of animals injected with pcDB, pcDC, pcDD or mix DNA did not change significantly after challenge with 2