关键词: 马立克氏病病毒, 细菌人工染色体, F因子

Abstract: [AIM] To construct the bacterial artificial chomosome containing the MDV1 genome, and further product fast and efficiently the recombinant virus of MDV1 by using BAC vector.[MAIN METHORD] Using the DNA recombinant techniques, the transferred vector containing E.coli F factor was constructed. The linearized vector was transfected into CEF which was infected by CVI988 virus. After three-cycle selection by MX-HAT, the infected CEF was stained with X-gal. [MAIN RESULT] The sequence analysis shows that the LoxP site synthesized in vitro was right,and the sequences of the homologous recombinant left arm and right arm are right. The two arms were ligated with the reversed oritation.The positive plasmid ,in which two LoxP sites had the same orientation, was identified by SphⅠand NheⅠ,named pUS-BGS. This linearized vector by NheⅠ was transfected into CEF which was infected by CVI988 virus. After three-cycle selection by MX-HAT, the infected CEF was stained with X-gal，blue plagues were appeared. [MAIN CONCLUSION] This research first reports that the transferred vector containing the E.coli F factor and two LoxP sites which had the same orientation, was constructed. The result shows that this vector is efficient for the selection of recombinant MDV1. So this gives the basis for construction of the bacterial artificial chromosome of MDV1.

Key words: Marek’s disease virus, bacterial artificial chromosome, F factor