Kisling U, Zlobin A Iu, Khil'ko S N
Vopr Virusol. 1981 Mar-Apr(2):210-5.
A system of cell-free synthesis from rabbit reticulocytes is described. A method of repeated bloodlettings for induction of reticulocytosis was used. The lysate of washed reticulocytes was treated with micrococcus nuclease enabling effective inhibition of synthesis in endogenous templates. The translating activity is manifested most completely in the presence of the following compounds: ATP, GTP, ATP-regenerating system, SH-compounds, polyamines, tRNA. This system must be optimized in K+ and Mg2+ concentrations for different templates. The system is tolerant to high concentrations of rRNA which allows the utilization of total RNA preparations in the experiments. This system may be satisfactorily used for translation of mRNA of different origins.
本文描述了一种来自兔网织红细胞的无细胞合成系统。采用了一种通过反复放血诱导网织红细胞增多的方法。洗涤后的网织红细胞裂解物用微球菌核酸酶处理,从而有效抑制内源性模板的合成。在以下化合物存在时,翻译活性表现得最为完全:ATP、GTP、ATP再生系统、含巯基化合物、多胺、tRNA。对于不同的模板,该系统必须在K⁺和Mg²⁺浓度方面进行优化。该系统耐受高浓度的rRNA,这使得在实验中可以使用总RNA制剂。该系统可令人满意地用于翻译不同来源的mRNA。