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本文采用 SDS-氯仿-苯酚法提取人胎肝总 RNA,每克胎肝组织可得总 RNA870～1060μg。用 Oligo(dT)纤维素亲合层析法分离出 Poly(A)~+mRNA,其得量为40～80μg/g 胎肝组织。将人胎肝来源的 Poly(A)~+mRNA 注射到非洲爪蟾卵母细胞内,经体外19℃孵育48h 后,在细胞提取液中可以检出刺激小鼠 CFU-S 增殖的刺激物,这表明人胎肝来源的 Poly(A)~+mRNA 中含有编码 CFU-S 增殖刺激物的 mRNA,并能在非洲爪蟾卵母细胞内翻译表达为活性蛋白质。
In this study, human fetal liver total RNA was extracted by SDS-chloroform-phenol method, and total RNA was 870 to 1060 μg per gram of fetal liver tissue. Poly(A)~+mRNA was isolated by Oligo(dT) cellulose affinity chromatography, and its yield was 40-80 μg/g fetal liver tissue. Human fetal liver-derived Poly(A)~+ mRNA was injected into Xenopus laevis oocytes and incubated at 19[deg.] C. for 48 h in vitro. Stimulators of CFU-S proliferation in mice were detected in cell extracts. This indicates that human fetal liver-derived poly(A)~+ mRNA contains mRNA encoding a CFU-S proliferation stimulator and can be translated into an active protein in Xenopus oocytes.