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以DNA病毒及正链RNA病毒为载体的研究已取得很大进展,而以负链RNA病毒为载体的研究直到近几年才获得较大突破.在以前的研究中,作者从已克隆的狂犬病病毒(RV)全长.cDNA获得了具有感染性的活RV,这为RV作为基因工程载体铺平了道路.在本研究中,作者以克隆的SAD B19株全长cDNA为基础,在RV基因组的G和(?)基因间插入一段N/P基因边界序列(含转录终止子及新的转录起始信号).然后将氯霉素乙酰转移酶(CAT)基因克隆在此序列下游,或者用以上序列(N/P基因边界序列及CAT全基因)替换(?)基因,从而获得两株克隆了外源基因的重组RV cDNA pSADXCAT及pSADXCAT.用表达RV N、P及L蛋白的
Research on DNA viruses and positive strand RNA viruses has made great progress, while negative strand RNA viruses have been the carriers for which breakthroughs have not been achieved until recent years.In previous studies, The full-length virus (RV) .cDNA obtained a viable live RV that paved the way for RV to function as a genetic engineering vector.In this study, based on the full-length cDNA of the cloned SAD B19 strain, (Including the transcription terminator and a new transcription initiation signal) between the G and (?) Genes, and then cloned the chloramphenicol acetyltransferase (CAT) gene downstream of this sequence or by using The above sequences (N / P gene border sequence and CAT whole gene) were substituted for the (?) Gene to obtain two recombinant RV cDNAs pSADXCAT and pSADXCAT with the exogenous genes cloned. Using the expression of RV N, P and L proteins