New rhinoviral DNA with mutation(s) in protease 2A region - and corresponding oligo:nucleotide(s) and oligopeptide(s), useful in test systems and as therapeutic competitive inhibitors

摘要

New DNA molecule (I) encoding for the VP1/P2A region of the rhinoviral system has, in the P2A region, non-wild type unique signal sequences for restriction enzymes. Also new are (1) oligonucleotides (II) encoding for a modified VP1/P2A region in which the modification is pref. present at cleavage sites or in the cleavage region of P2A, and esp. modified cis activity; (2) test systems contg. (I) in a suitable expression system; (3) oligopeptides (III) encoded by (II), and (4) oligopeptides (IIIa) split by P2A in trans. The rhinovirus is of serotypes HRV2 or HRV89 and the signal sequences for the enzymes ZmaI, SmaI, XmaIII, AvaI, NaeI, MluI, EagI, BglII or SnaBI are recognisable. In (II) the modification results in destruction of the P2A active centres. USE/ADVANTAGE - The test systems allow examination of cis and trans activity, and expression systems contg. (I), which encode for the same amino acid sequence as native viral DNA and differ only in restriction enzyme pattern, can be used to define the smallest region of HRV which retains proteolytic activity. (III) are useful as competitive inhibitors of trans activity so can be used therapeutically or prophylactically.