MRNA-directed synthesis of catalytically active mouse beta-glucuronidase in Xenopus oocytes.
MRNA-directed synthesis of catalytically active mouse beta-glucuronidase in Xenopus oocytes.
复制标题
非洲爪蟾卵母细胞中催化活性小鼠 β-葡萄糖醛酸酶的 mRNA 指导合成。
DOI：
10.1073/pnas.74.10.4462
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发表时间：
1977
影响因子：
11.1
通讯作者：
K. Paigen
中科院分区：
文献类型：
--
作者：
C. Labarca;K. Paigen
当向爪蟾卵母细胞注射富含含poly（A）的mRNA序列的小鼠RNA时，形成具有催化活性的小鼠β-葡萄糖醛酸苷酶（β-D-葡糖苷酸葡糖醛酸水解酶，EC 3.2.1.31）。对于来自雄激素诱导的肾脏的RNA，翻译效率与内源性爪蟾信使的翻译效率相当，并且翻译的保真度很高。通过形成催化活性产物检测葡萄糖醛酸苷酶信使比通过掺入同位素标记的氨基酸检测灵敏几个数量级。该系统不仅提供了一种检测基因表达调节的灵敏技术，还提供了一个研究溶酶体酶翻译后多肽加工调节的机会。
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.