摘要
构建腺病毒穿梭载体pAd RSV ,并将p3 8MAPK (mitogen activatedproteinkinase)的上游激酶MKK6(mitogen activatedproteinkinasekinase 6)及其持续激活和无活性的突变体基因亚克隆到该穿梭载体 .通过与腺病毒DNA(pJM17)在能够表达E1的HEK 2 93细胞同源重组生成了能够表达MKK6信号分子的重组腺病毒 .PCR结果表明 ,这些重组腺病毒DNA的插入片段大小是正确的 .而且 ,通过感染COS 7细胞 ,用免疫激酶活性测定证实这些重组的腺病毒能够表达具有功能的蛋白质 .
The construction of pAd/RSV shuttle vector for adenovirus was performed.Mitogen activated protein kinase kinase 6 (MKK6),an upstream kinase of p38 mitogen activated protein kinase (MAPK),and its constitutive and inactive mutants were subcloned into the shuttle vector.The recombinant adenoviruses expressing proteins of MKK6 or its mutants were produced by homologous recombination of shuttle vectors and adenovirus DNA pJM17 in HEK 293 cells expressing E1 gene.The PCR results showed that the insertions of the recombinant adenoviruses were of the right size.It was proved by immuno kinase assay that these recombinant adenoviruses expressed functional proteins in COS 7 cells with the infection of these recombinant viruses.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2001年第5期595-601,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
"8 6 3"生物技术领域青年科学基金
国家自然科学基金 ( 3 980 0 0 6 0 )
