Grnr.
161 ( 1995) 297-X
‘0 1995 Elsevier Science B.V. All rights reserved.
297
0378-I 119~95~$09.50
GENE08973
Cloning and sequencing of the F gene of live attenuated Urabe Am9 mumps virus (Recombinant
DNA; vaccine strain; wild-type MpsV virus; cDNA homologies)
M. Grazia Cusi, Silvia Bianchi, Marcello
Received by J.A. Engler: I8 January
Valassina, Laura Santini and Pier E. Valensin
1995; Revised/Accepted:
7 February.;8
February
1995; Received at publishers:
7 April 1995
SUMMARY
A cDNA clone encoding the entire F gene of the live attenuated mumps virus (MpsV) strain, Urabe Am9, has been isolated and sequenced. The F gene sequence shows significant homology with the one reported for the wild-type MpsV Miyahara strain.
Several strains of attenuated mumps virus (MpsV) have been developed for use in vaccines. The Urabe Am9 strain of attenuated live MpsV (MpsV-Va) is produced either in the amnion of embryonated hen’s eggs or in chick embryo cell cultures (Yamanishi et al., 1970). MpsV-Va cDNA (Sclavo, Siena, Italy) was obtained by screening a library from mRNA of virally-infected cells with a synthetic probe (5’-GTGGAAGTGTAACGTGTCTAT), corresponding to a fragment of the Miyahara strain (MpsV-M) F gene. Comparison with the reported sequences of RW (Waxham at al., 1987) SBL-1 (Elliott et al., 1989) and MpsV-M (Takeuchi et al., 1989) strains revealed that the MpsV-Ur strain showed 69 (homology approx. 96%) 101 (homology approx. 94%) and 23 (homology approx. 98%) nt differences corresponding to 12, 24 and 3 aa differences, respectively. It appears that the MpsV-Ur strain is most closely related to the Mpsv-M strain, since the three aa substitutions
Correspondenc~r
to: Dr. M.G. Cusi, Department
of Molecular
Biology,
Section of Microbiology, University of Siena, Via Laterina, 8. 53100 Siena. Italy. Tel. (39-577) 280-903: Fax (39-577) 42011: e-mail: Cusi(Zirjumst.it Abbreviations:
aa,
Miyahara strain nucleotide(s). Ssnl
0378-l
amino
acid(s);
of MpsV; MpsV-Ur.
119(95100255-3
MpsV. Urabe
mumps
virus;
Am9 strain
MpsV-M.
of MpsV; nt.
(Ala3+Val; Ser” +Gly; Ileho -+Thr) do not represent critical changes (Fig. 1).
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298
600 104 750 234 900 284
,200 384 L350 434 1500 484 PSjCT~~GTAGCAGCC~~~~A~~-~A~~-TA~T~~~-TT~A~T~T
CRWLlrOMTCAACTTC~~TA~T~~T~~T~~~~~ ....AIIVAALVLSILSIIISLLPCCWAYIATKElRRINPKTNH~*T~SSS"~~ .............................................................................................................................................................................
1650 534 L713 538
Fig. 1. Nucleotide sequence of MpsV F gene and the deduced aa sequence. The presumptive signal sequence of the fusion protein of MpsV-Ur strain is underlined. The likely transmembrane anchoring region is highlighted by a dotted line under the sequence. EMBL accession No. X82887 MVIRFPRO.