Richardson W D, Carter B J, Westphal H
Proc Natl Acad Sci U S A. 1980 Feb;77(2):931-5. doi: 10.1073/pnas.77.2.931.
Adenovirus type 2 mRNAs were injected via glass capillaries into Vero cells, a line of African green monkey kidney cells permissive for adenovirus growth. Polypeptides synthesized after injection were labeled with 35S-labeled amino acids, precipitated with antiviral sera, and analyzed by polyacrylamide gel electrophoresis. Both early and late viral mRNAs give rise to authentic polypeptides. The in vivo translation of mRNAs can be measured as late as 24 hr after injection. The ability to analyze the protein products of microinjected mRNAs directly should greatly extend the applications of the procedure. Vero cells injected with early mRNA from adenovirus type 2 support the growth of adenovirus-associated virus, a defective virus that is dependent on adenovirus helper functions. This result demonstrates that a measurable biological activity, the ability to overcome the defectiveness of adenovirus-associated virus, resides in early adenovirus mRNA.
将2型腺病毒信使核糖核酸(mRNA)通过玻璃毛细管注入非洲绿猴肾细胞系Vero细胞中,该细胞系允许腺病毒生长。注射后合成的多肽用35S标记的氨基酸进行标记,用抗病毒血清沉淀,并用聚丙烯酰胺凝胶电泳进行分析。早期和晚期病毒mRNA均可产生真实的多肽。mRNA的体内翻译在注射后24小时仍可检测到。直接分析显微注射mRNA的蛋白质产物的能力应能大大扩展该方法的应用。注射了2型腺病毒早期mRNA的Vero细胞支持腺病毒相关病毒的生长,腺病毒相关病毒是一种依赖腺病毒辅助功能的缺陷病毒。这一结果表明,早期腺病毒mRNA具有一种可测量的生物活性,即克服腺病毒相关病毒缺陷的能力。