Samols D, MacIntyre S S, Kushner I
Biochem J. 1985 May 1;227(3):759-65. doi: 10.1042/bj2270759.
C-reactive protein (CRP) mRNA was assayed by cell-free translation of poly(A)-containing liver RNA isolated both from rabbits stimulated to undergo the acute-phase response and from unstimulated control rabbits. No CRP-related translation products were identified until the denaturant methylmercury hydroxide (CH3HgOH) was added to the RNA before cell-free translation. In the presence of the denaturant, a 24000-Da translation product was synthesized which was immunochemically identifiable as the CRP primary translation product. It is likely that rabbit CRP mRNA can form a stable intramolecular duplex which interferes with its translatability in vitro. The 24000-Da CH3HgOH-facilitated cell-free translation product was not detected in poly(A)-containing liver RNA from unstimulated animals, indicating that the concentration of translatable CRP mRNA was dramatically induced during the acute-phase response. On the basis of absorption experiments, the 24000-Da CRP primary translation product was immunochemically more closely related to denatured CRP than to native CRP.
通过对从经历急性期反应的兔子以及未受刺激的对照兔子中分离出的含聚腺苷酸(poly(A))的肝脏RNA进行无细胞翻译,来检测C反应蛋白(CRP)mRNA。在无细胞翻译前,向RNA中加入变性剂氢氧化甲基汞(CH3HgOH)之前,未鉴定出与CRP相关的翻译产物。在变性剂存在的情况下,合成了一种24000道尔顿的翻译产物,经免疫化学鉴定为CRP初级翻译产物。兔CRP mRNA很可能形成了一种稳定的分子内双链体,从而在体外干扰其翻译能力。在未受刺激动物的含poly(A)的肝脏RNA中未检测到24000道尔顿的CH3HgOH促进的无细胞翻译产物,这表明在急性期反应期间可翻译的CRP mRNA浓度显著增加。基于吸收实验,24000道尔顿的CRP初级翻译产物在免疫化学上与变性CRP的关系比与天然CRP的关系更密切。