Cloning the double-stranded RNA genes of reovirus: sequence of the cloned S2 gene.
Cloning the double-stranded RNA genes of reovirus: sequence of the cloned S2 gene.
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DOI：
10.1073/pnas.79.24.7644
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发表时间：
1982-12
影响因子：
11.1
通讯作者：
L. Cashdollar;J. Esparza;G. Hudson;R. Chmelo;P. Lee;W. Joklik
中科院分区：
文献类型：
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作者：
L. Cashdollar;J. Esparza;G. Hudson;R. Chmelo;P. Lee;W. Joklik
呼肠孤病毒血清型3的Dearing株的基因由双链RNA组成，通过用poly（A）给每个基因的两条链加尾，用逆转录酶转录它们，使同源的正和负cDNA链自杂交，将它们与大肠杆菌DNA聚合酶I孵育以确保它们是完整的，并通过标准方法克隆双链cDNA分子。克隆的S2基因的序列已经确定。末端的序列与天然双链RNA基因末端的序列完全相同。该基因长1，329个核苷酸，具有一个单一的长开放阅读框，其起始于第一个起始密码子（残基19）并延伸331个密码子，足以编码与已知的S2基因产物，蛋白质sigma 2，主要呼肠孤病毒核心组分（Mr，38，000）相同大小的蛋白质。85个密码子的第二个开放阅读框在不同的阶段开始于接近第一个结束的地方。从这个阅读框翻译的蛋白质是未知的。
The genes of the Dearing strain of reovirus serotype 3, which consist of double-stranded RNA, have been cloned into pBR322 by tailing both strands of each gene with poly(A), transcribing them with reverse transcriptase, self-hybridizing the cognate plus and minus cDNA strands, incubating them with Escherichia coli DNA polymerase I to ensure that they are complete, and cloning the double-stranded cDNA molecules by standard procedures. The sequence of the cloned S2 gene has been determined. The sequence at the termini are exactly the same as those at the ends of the native double-stranded RNA gene. The gene is 1,329 nucleotides long and possesses a single long open reading frame that starts at the first initiation codon (residue 19) and extends for 331 codons, sufficient to encode a protein of the same size as the known S2 gene product, protein sigma 2, a major reovirus core component (Mr, 38,000). A second open reading frame of 85 codons, in a different phase, starts close to where the first ends. The protein translated from this reading frame is unknown.