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Cloning and sequencing of the gene encoding the spike protein of the coronavirus infectious bronchitis virus

Cloning and sequencing of the gene encoding the spike protein of the coronavirus infectious bronchitis virus

Journal of General Virology 66(4): 719-726

RNA sequences encoding the surface projection (spike) of the coronavirus infectious bronchitis virus, strain Beaudette, were cloned into pBR322 using complementary DNA primed with a specific oligonucleotide. A 5.3 kilobase viral insert in the clone pMB179 was identified. The region of this clone coding for the spike gene was sequenced by the chain termination method, and the 1st report of DNA sequence data for a coronavirus spike protein, the protein which forms the characteristic corona after which the group is named, is presented here. The amino acid sequence of the primary translation product, deduced from the DNA sequence, predicts a polypeptide of 1162 amino acids with a MW of 127,006. This has many interesting features which confirm and extend knowledge of this recently characterized membrane glycoprotein. The polypeptide is subsequently cleaved to S1 and S2, and partial amino acid analysis of the amino-terminus of the S1 polypeptide was employed to locate the position of this terminus of S1 within the large open reading frame. The amino acid analysis also reveals the presence of an 18 amino acid putative signal sequence on the primary translation product which is not present on the mature S1 polypeptide.

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