Kidson S H, Fabian B C
Biochim Biophys Acta. 1985 Jan 29;824(1):40-4. doi: 10.1016/0167-4781(85)90027-2.
We present a method whereby some mRNAs which code for enzymes can be isolated by affinity chromatography of newly synthesized polypeptides bound to their polysome complexes. Using this method we have isolated tyrosinase-mRNA from Xenopus laevis oocytes and have analysed the translation products from the RNAs thus obtained. In vitro translation reveals the presence of two mRNAs coding for polypeptides of molecular weights of 20 000 and 32 000, respectively. The larger molecule corresponds to the molecular weight of nascent tyrosinase. Furthermore, microinjection of the mRNA into Xenopus oocytes results in the synthesis of active tyrosinase. Since this isolation method is dependent on the ability of nascent enzymes to bind to their substrate analogue, it is thought that this approach may be appropriate for obtaining mRNAs coding for other enzymes.
我们提出了一种方法，通过该方法可以通过对与其多核糖体复合物结合的新合成多肽进行亲和层析来分离一些编码酶的mRNA。使用这种方法，我们从非洲爪蟾卵母细胞中分离出了酪氨酸酶mRNA，并分析了由此获得的RNA的翻译产物。体外翻译显示存在两种分别编码分子量为20000和32000的多肽的mRNA。较大的分子对应于新生酪氨酸酶的分子量。此外，将该mRNA显微注射到非洲爪蟾卵母细胞中会导致活性酪氨酸酶的合成。由于这种分离方法依赖于新生酶与其底物类似物结合的能力，因此认为这种方法可能适用于获得编码其他酶的mRNA。
