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Turku Immunology Centre and Department of Virology, University of Turku, Kiinamyllynkatu 13, FIN-20520 Turku, Finland
We have determined the nucleotide sequences of the capsid, E3, E2 and 6K genes of the avirulent Semliki Forest virus variant A774 (SFV A7). The sequence analysis revealed a nucleotide identity of 98% for capsid, 98% for E3, 97% for E2 and 98% for 6K genes, as compared with the prototype SFV strain L10. At the protein level, the capsid and E3 polypeptides of SFV A7 both exhibited two amino acid substitutions, whereas point mutations in the 6K gene did not alter the amino acid sequence. In the E2 gene of SFV A7, seven of the 34 point mutations led to an amino acid difference as compared with the L10 strain. Replacement of the E2 glycoprotein gene of the virulent SFV4 clone with the corresponding region of SFV A7 resulted in a new plasmid construct, pME2, that gave rise to infectious virus CME2. CME2 and SFV4 replicated similarly in an immortalized mouse brain cell line (MBA 13). Intraperitoneal injection of 106 p.f.u. of CME2 into 4- to 6- week-old BALB/c mice caused mild clinical signs in some mice, whereas the majority of the infected animals remained asymptomatic, similar to infection with the avirulent SFV A7. In contrast, infection with the parental SFV4, a derivative of the virulent L10 strain, was lethal in 80% of mice. Virus titres in blood and brain tissue specimens of BALB/c mice were similar after infection with CME2 or A7 viruses. The results suggest that amino acid differences in the E2 glycoprotein individually or in concert cause the attenuation of CME2.
* Author for correspondence. Fax +358 21 6337000. e-mail ahinkkanen@finabo.abo.fi
Received 19 May 1994;
accepted 26 August 1994.
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