Cloning and sequencing of the F gene of live attenuated Urabe Am9 mumps virus

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.