Development of paramyxovirus vector

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Paramyxoviruses are negative-stranded RNA virus and have interesting characteristic of cell tropisms and replication modes and may be usuful for vector for introducing exogeneous genes into cells. For this porpose, we intended to make mumps virus rescue system from cDNA.As a initial step, we constructed a minigenome containing reporter cat (chloramphenicol acetyl transferase) gene flanked by the mumps virus 5' and 3' end Sequences. When a minigenome was transfected into the cells expressing the L,NP and P protein from cDNAs, unfortunately, no CAT activity was detected in the cell lysates indicating no replication of the minigenome. We determined the nucleotide sequences of the NP,P,L plasmids and also the nuclotide sequences of the RT-PCR fragments synthesized from the mumps virus genomic RNA and any problem could not be detected. As it was reported that the cat minigenome system of some paramixoviruses did not work for unknown reason, this may be also the case for mumps virus. During the course of experiments, we found tha mumps virus SH gene is not essential for virus growth and SH gene unit may be deleted and replaced with other genes of interst, and we established cell lines stably expressing the mumps virus F or HN proteins and these cell may be useful to recover the mumps virus without the F or HN genes. To identify the reasons for inability of the replication of the mumps minigenome, we tried the measles virus rescue system developed by Dr.Billeter. As we were able to recover infectious measles virus from the measles full-length cDNA and detect CAT activity from the cell lysate transfected with measles minigenome, we do not believe that basic techniques we used are problem. We are now constructing complete cDNA of the mumps virus genome.