Shanks M, Dessens J T, Lomonossoff G P
Department of Virus Research, John Innes Centre, Norwich, UK.
J Gen Virol. 1996 Sep;77 ( Pt 9):2365-9. doi: 10.1099/0022-1317-77-9-2365.
To investigate the specificity of comoviral 24 kDa ('24K') proteinases, a full-length cDNA copy of red clover mottle virus (RCMV) RNA 1 has been cloned downstream of a T7 promoter. Translation in rabbit reticulocyte lysates of in vitro transcripts from this clone resulted in the synthesis of a 200K protein which was processed in a manner similar to that of the equivalent protein from cowpea mosaic virus (CPMV). Full-length cDNA clones of the RNA 1 molecules of RCMV and CPMV were used to create hybrid RNA 1 molecules. RNA transcribed in vitro from these hybrids was translated in vitro and the ability of the 24K proteinase from one comovirus to cleave the 32K/170K processing site from the other assessed. The results of the experiments show that the 24K proteinases are virus-specific in cis.
为了研究伴生病毒24千道尔顿(“24K”)蛋白酶的特异性,已将红三叶草斑驳病毒(RCMV)RNA 1的全长cDNA拷贝克隆到T7启动子下游。该克隆体外转录本在兔网织红细胞裂解物中的翻译产生了一种200K蛋白,其加工方式与豇豆花叶病毒(CPMV)的等效蛋白相似。利用RCMV和CPMV的RNA 1分子的全长cDNA克隆构建了杂交RNA 1分子。这些杂交体体外转录的RNA在体外进行翻译,并评估了一种伴生病毒的24K蛋白酶切割另一种伴生病毒32K/170K加工位点的能力。实验结果表明,24K蛋白酶在顺式作用中具有病毒特异性。