Roy P, Purdy M A, Petre J, Rao C D
Prog Clin Biol Res. 1985;178:363-70.
This is the 1st report of molecular cloning of bluetongue virus (BTV) gene using DNA recombinant techniques. A number of complete and overlapping clones of BTV genes have been cloned into pBR322 by standard procedures with some modification. The full length clones were confirmed by matching the terminal cDNA nucleotide sequences with those previously determined for the termini of double-stranded genomic RNA (Rao et al., 1983). The complete sequence of the cDNA clone of RNA segment 3 of US serotype 17 as determined by the method of Maxam and Gilbert (1980) has been presented here. The clone is 2,772 nucleotides (1.78 X 10(6) daltons) long excluding the 3' polyadenylic acid sequence and has an open reading frame which encodes a protein of some 901 amino acids (103,412 daltons).
这是关于使用DNA重组技术对蓝舌病毒(BTV)基因进行分子克隆的首次报道。通过一些改进的标准程序,已将多个完整且重叠的BTV基因克隆到pBR322中。通过将末端cDNA核苷酸序列与先前确定的双链基因组RNA末端序列进行比对(Rao等人,1983年),确认了全长克隆。本文展示了通过Maxam和Gilbert(1980年)方法确定的美国17型血清型RNA片段3的cDNA克隆的完整序列。该克隆长2772个核苷酸(1.78×10⁶道尔顿),不包括3'聚腺苷酸序列,并且具有一个开放阅读框,编码约901个氨基酸的蛋白质(103412道尔顿)。