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兔α和β珠蛋白mRNA 5'末端的核苷酸序列。

Nucleotide sequences at the 5'termini of rabbit alpha and beta globin mRNA.

作者信息

Lockhard R E, Rajbhandary U L

出版信息

Cell. 1976 Dec;9(4 PT 2):747-60. doi: 10.1016/0092-8674(76)90138-0.

Abstract

The nucleotide sequences at the 5' termini of rabbit alpha and beta globin mRNAs have been determined. Periodate oxidation of globin mRNA followed by reduction with 3H-sodium borohydride and subsequent analysis of the 3H-labeled mRNA reveals the presence of the "cap" structure m7G5'ppp- blocking the 5' terminus. After periodate oxidation, beta elimination, and phosphomonoesterase treatment to remove the m7G5'ppp- "cap," the 5' end of globin mRNA was labeled with 32P using gamma-32P-ATP and T4 polynucleotide kinase. The 5'-32P-labeled alpha and beta globin mRNAs were then resolved from each other by polyacrylamide gel electrophoresis under denaturing conditions and sequenced separately. The 5' terminal nucleotide sequences determined are: alpha--m7G5'ppp5m6AmC(m)ACUUCUGG- BETA--M7G5'ppp5m6AmC(m) ACUUGCUUUUGACACAA Besides the m7G "cap" structure, the two sequences are identical for the first six nucleotides and then diverge. No initiator codon is present within the first ten nucleotides from the 5' end of the alpha globin mRNA, and the first nineteen nucleotides from the 5' end of beta globin mRNA.

摘要

已确定兔α和β珠蛋白mRNA 5'末端的核苷酸序列。对珠蛋白mRNA进行高碘酸盐氧化,然后用3H-硼氢化钠还原,随后对3H标记的mRNA进行分析,结果显示存在封闭5'末端的“帽”结构m7G5'ppp-。在高碘酸盐氧化、β消除和磷酸单酯酶处理以去除m7G5'ppp-“帽”之后,使用γ-32P-ATP和T4多核苷酸激酶用32P标记珠蛋白mRNA的5'末端。然后在变性条件下通过聚丙烯酰胺凝胶电泳将5'-32P标记的α和β珠蛋白mRNA彼此分离并分别测序。所确定的5'末端核苷酸序列如下:α——m7G5'ppp5m6AmC(m)ACUUCUGG-β——M7G5'ppp5m6AmC(m)ACUUGCUUUUGACACAA除了m7G“帽”结构外,这两个序列的前六个核苷酸相同,然后不同。在α珠蛋白mRNA 5'末端的前十个核苷酸内以及β珠蛋白mRNA 5'末端的前十九个核苷酸内均不存在起始密码子。

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