Maniatis G M, Ramirez F, Cann A, Marks P A, Bank A
J Clin Invest. 1976 Dec;58(6):1419-27. doi: 10.1172/JCI108597.
Human globin messenger RNA (mRNA) prepared from erythroid cells of patients with sickle cell anemia has been translated in Xenopus laevis oocytes. Addition of hemin to the injected mRNA causes total globin synthesis to increase and the ratio of alpha- to betas-globin synthesis (alpha/betas ratio) to approach unity. To determine the effect of the length of the poly-(A) segment on human globin mRNA stability, 10 S globin mRNA was fractionated into poly-(A)-poor fractions by oligo (dT)-cellulose column chromatography. When oocytes are injected with each of these fractions, translation of the poly-(A)-rich globin mRNA is sustained for a longer period than that of the poly-(A)-poor mRNA. Regardless of the mRNA fraction injected, the alpha/betas ratio of the synthesized globin decreases as the injected oocytes are incubated for longer periods. The results indicate that in frog oocytes poly-(A)-rich mRNA has greater translational stability than poly-(A)-poor mRNA, AND beta-mRNA has greater stability than alpha-mRNA with comparable poly-(A) content.
从镰状细胞贫血患者的红细胞中制备的人珠蛋白信使核糖核酸(mRNA)已在非洲爪蟾卵母细胞中进行翻译。向注射的mRNA中添加血红素会使总珠蛋白合成增加,且α珠蛋白与β珠蛋白合成的比例(α/β比例)接近1。为了确定聚腺苷酸(poly - (A))片段长度对人珠蛋白mRNA稳定性的影响,通过寡聚(dT)-纤维素柱色谱法将10S珠蛋白mRNA分离成富含poly - (A)的部分和poly - (A)含量低的部分。当向卵母细胞注射这些部分中的每一部分时,富含poly - (A)的珠蛋白mRNA的翻译持续时间比poly - (A)含量低的mRNA更长。无论注射的是哪种mRNA部分,随着注射的卵母细胞孵育时间延长,合成的珠蛋白的α/β比例都会降低。结果表明,在蛙卵母细胞中,富含poly - (A)的mRNA比poly - (A)含量低的mRNA具有更高的翻译稳定性,并且在poly - (A)含量相当的情况下,β - mRNA比α - mRNA具有更高的稳定性。