Heterogeneity of the 5' terminus of hen ovalbumin messenger ribonucleic acid.
Heterogeneity of the 5' terminus of hen ovalbumin messenger ribonucleic acid.
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母鸡卵清蛋白信使核糖核酸 5 末端的异质性。
DOI：
10.1093/nar/9.7.1657
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发表时间：
1981
影响因子：
14.9
通讯作者：
Rhoads,RE
中科院分区：
文献类型：
--
作者：
Malek,LT;Eschenfeldt,WH;Munns,TW;Rhoads,RE
用一种新的标记方法对鸡卵清蛋白mRNA的5′-末端序列进行了研究。通过与偶联至纤维素的互补DNA杂交来纯化卵清蛋白mRNA。由此纯化的mRNA通过与含有编码伴清蛋白和卵类粘蛋白的信使序列的质粒DNA杂交显示纯度为97.9%，伴清蛋白和卵类粘蛋白是输卵管中接下来的两个最丰富的信使。经RNase T1和碱性磷酸酶酶切后，通过与抗m7 G-Sepharose结合来选择5′端加帽的寡核苷酸。然后用RNA连接酶和[5′-32 P]pCp标记，通过二维凝胶电泳分离，并通过碱基特异性核糖核酸酶部分消化进行测序。鉴定了一组嵌套的三种加帽寡核苷酸。它们的结构和相对丰度分别为：m7 GpppAUACAG，3%，m7GpppAACAG61%，m7GpppGUAACAG36%。
The 5′-terminal sequence of hen ovalbumin mRNA was investigated using a novel labeling method. Ovalbumin mRNA was purified by hybridization to complementary DNA coupled to cellulose. The mRNA thus purified was shown to be 97.9% pure by hybridization with plasmid DNA containing sequences to the messengers coding for conalbumin and ovomucoid, the next two most abundant messengers of oviduct. After digestion with RNase T1and alkaline phosphatase, 5′-terminal capped oligonucleotides were selected by binding to anti-m7G-Sepharose. These were then labeled using RNA ligase and [5′-32P]pCp, separated by two-dimensional gel electrophoresis, and sequenced by partial digestion with base-specific ribonucleases. A nested set of three capped oligonucleotides was identified. Their structures and relative abundances were m7GpppAUACAG, 3%; m7GpppACAUACAG, 61% and m7GpppGUACAUACAG, 36%.