CLONING AND SEQUENCING OF THE GENE ENCODING THE SPIKE PROTEIN OF THE CORONAVIRUS-IBV
CLONING AND SEQUENCING OF THE GENE ENCODING THE SPIKE PROTEIN OF THE CORONAVIRUS-IBV
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DOI：
10.1099/0022-1317-66-4-719
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发表时间：
1985-01-01
影响因子：
3.8
通讯作者：
BROWN, TDK
中科院分区：
文献类型：
--
作者：
BINNS, MM;BOURSNELL, MEG;BROWN, TDK
使用用特异性寡核苷酸引发的互补DNA，将编码冠状病毒传染性支气管炎病毒Beaudette株的表面突起（刺突）的RNA序列克隆到pBR 322中。鉴定了克隆pMB 179中的5.3内切酶病毒插入物。用链终止法对该克隆的刺突基因编码区进行了测序，并首次报告了冠状病毒刺突蛋白的DNA序列数据，该蛋白形成了该组命名的特征冠。从DNA序列推断的初级翻译产物的氨基酸序列预测具有127，006的MW的1162个氨基酸的多肽。这有许多有趣的功能，确认和扩展知识，这最近的特点膜糖蛋白。随后将多肽裂解为S1和S2，并对S1多肽的氨基末端进行部分氨基酸分析，以确定S1的该末端在大开放阅读框内的位置。氨基酸分析还揭示了在初级翻译产物上存在18个氨基酸的推定信号序列，其不存在于成熟S1多肽上。
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.