Labarca C, Paigen K
Proc Natl Acad Sci U S A. 1977 Oct;74(10):4462-5. doi: 10.1073/pnas.74.10.4462.
Catalytically active mouse beta-glucuronidase (beta-D-glucuronide glucuronosohydrolase, EC 3.2.1.31) is formed when Xenopus oocytes are injected with mouse RNA enriched for poly(A)-containing mRNA sequences. With the RNA from androgen-induced kidneys, the efficiency of translation is comparable to that of endogenous Xenopus messenger, and the fidelity of translation is high. Detection of glucuronidase messenger by formation of a catalytically active product is several orders of magnitude more sensitive than detection by incorporation of isotopically labeled amino acids. As well as providing a sensitive technique for examining the regulation of gene expression, the system makes available an opportunity to study the regulation of post-translational polypeptide processing of a lysosomal enzyme.
当向非洲爪蟾卵母细胞注射富含聚腺苷酸(poly(A))的mRNA序列的小鼠RNA时,可形成具有催化活性的小鼠β-葡萄糖醛酸酶(β-D-葡萄糖醛酸葡萄糖醛酸水解酶,EC 3.2.1.31)。使用来自雄激素诱导的肾脏的RNA,翻译效率与内源性非洲爪蟾信使RNA相当,且翻译保真度高。通过形成具有催化活性的产物来检测葡萄糖醛酸酶信使RNA比通过掺入同位素标记的氨基酸进行检测要灵敏几个数量级。该系统不仅提供了一种检测基因表达调控的灵敏技术,还为研究溶酶体酶翻译后多肽加工的调控提供了机会。