Method for improving genetic stability of foreign insert nucleotide sequence in recombinant poliovirus

DETAILED DESCRIPTION To be free from shortcomings of the poliovirus vector systems, in particular, genetic instability of the foreign insert integrated into the recombinant polioviruses, the present inventors have examined potential factors governing genetic stability of foreign insert by constructing and exploiting many different recombinant polioviruses, which contain a series of different original or sequence-adjusted foreign inserts.
From these experiments, we have accomplished present invention that i) the insert genetic stability is strongly associated with the G/C contents and its distribution patterns within the size limitation, and ii) the insert genetic stability can be markedly enhanced by increasing the G/C contents of the foreign insert.
Accordingly, it is an object of this invention to provide a method for improving a genetic stability of a foreign insert nucleotide sequence in a recombinant single-stranded RNA virus vector.
It is another object of this invention to provide a method for constructing a recombinant single-stranded RNA virus containing a foreign insert nucleotide sequence with improved genetic stability.
It is still another object of this invention to provide a recombinant single-stranded RNA virus comprising a foreign insert nucleotide sequence with improved genetic stability.
It is further object of this invention to provide a recombinant poliovirus comprising a foreign insert nucleotide sequence with improved genetic stability.
It is still further object of this invention to provide a method for construction of sequence-adjusted or artificially synthesized foreign inserts using template/ligation-free PCR method.
It is another object of this invention to provide a vaccine composition comprising a recombinant single-stranded RNA virus.
Other objects and advantages of the present invention will become apparent from the detailed description to follow taken in conjugation with the appended claims and drawings.