【摘要】 以A型口蹄疫病毒(FMDV)AV88(L)和XJ99株的P12X基因和AV88(L)3C基因的阳性克隆质粒为模板,用PCR扩增各基因片段,酶切后分步定向亚克隆至逆转录病毒表达载体pBABE-puro上经PCR、酶切鉴定及测序。结果表明,获得了2个含不同RGD基序的A型FMDV衣壳蛋白和蛋白酶基因的重组逆转录病毒表达载体pBABE-AV88(L)P1 2X3C和pBABE-XJ99 P1 2X3C,为研究安全、高效、低成本的FMDV空衣壳亚单位疫苗奠定了基础。更多还原

【Abstract】 By using two-step cloning strategy,the fragment P1 2X3C of foot and mouth disease virus serotype A was obtained and two recombinant retrovirus expression vectors pBABE-AV88(L)P1 2X3C and pBABE XJ99 P1 2X3C were constructed.P1 2A and 3C genes with flanking restriction sites for endo- nuclease were amplified respectively by PCR,and then the P1 2X gene was digested by BamHⅠand EcoRⅠ.The gene P1 2X was cloned into the pBABE expression vector which was digested by the same en- zyme.The recombinant plasmid was identified by PCR,digestion with BamHⅠ+EcoRⅠand sequen- cing.After the Pl 2X plasmid was constructed,the gene 3C flanked EcoRⅠwas cloned into the Pl 2X positive expression plasmid.The result indicated that the fragment P1 2X3C was cloned into the retrovirus expression plasmid correctly.更多还原