鸡马立克氏病基因工程二价苗的初步研究

Preliminary Study on the Genic Engineering Bivalent Vaccine of Chicken Marek's Disease

用磷酸钙沉淀法将MDⅠ型病毒gB基因插入到HVT病毒载体中制成马立克重组病毒DNA,再将重组DNA转染于鸡胚成纤维细胞上,形成的病毒蚀斑与HVT病毒蚀斑形态相似,经复制2～3代后,提纯病毒DNA,PCR扩增后电泳分析,凝胶上形成一条2.9kb左右的条带。将重组病毒负染后用电镜观察,病毒粒子形态与HVT病毒相似。结果表明,gB基因已经重组在二价病毒中,并能够稳定遗传。

The recombinant MD virus DNA was constructed by inducing the gB gene of MD 1 type virus into the HVT DNA vector through Calcium phosphate precipitation. The pathological plaques identical with those of HVT virus formed after transfecting the recombinant DNA in CEF cells. The DNA was purified, PCR amplified and electrophoresed after replicated 2 or 3 generations, a band about 2.9 kb was formed on the electrophoresis gel. The recombinant virus is identical with the HVT virus in shape under the electronic microscope after negative dyeing. The result shows that the gB gene was recombined into the virus and can inherit steadily.