摘要
鸡痘病毒含有动物病毒中最大的基因组.本研究以pUC18质粒作载体,对鸡痘病毒282E_4弱毒株基因组的EcoRI酶切片段进行了鸟枪法克隆及5.5kb EcoRI片段的定向克隆.根据β-半乳糖苷酶显色反应选择法的初步筛选和以[α-^(32)P-dATR]标记的鸡痘病毒DNA为探针进行的原位杂交、打点杂交以及Southem印迹杂交结果,证实已成功地克隆了鸡痘病毒的某些DNA片段.
Fowlpox virus (FPV) is the type species of the Avipoxvirus group of the poxviridae. It's a promising vector to express foreign genes. The EcoRI restriction fragments of 284E4 attenuated strain genome were cloned into pUC18 vector using the shotgun method, and the 5.5 kb EcoRI fragment was cloned The recombinant colonies were selected according to the phenotypes of inactivation of Lac Z gene and the inserts were further identified by in situ colony hybridization, dot hybridization with [α-32P-dATP] -labeled FPV DNA. The experiment has laid the foundation for further cloning of the nonessential genes and the promotor sequences of the FPV genome.
