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使用特异性引物和RNA测序的双脱氧核苷酸方法对免疫球蛋白mRNA进行完整测序。

Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing.

作者信息

Hamlyn P H, Gait M J, Milstein C

出版信息

Nucleic Acids Res. 1981 Sep 25;9(18):4485-94. doi: 10.1093/nar/9.18.4485.

Abstract

The complete sequence of the mouse immunoglobulin kappa light chain MOPC 21 messenger RNA has been determined using a chain termination method and chemically synthesised deoxyoligonucleotides to initiate the synthesis of a DNA molecule complementary to the mRNA template. Five such oligonucleotide primers have been used for the sequence analysis of this messenger RNA. The approach is excellent for comparative studies of mouse k-chain mRNAs because they can be made on impure mRNA preparations. The MOPC 21 light chain mRNA is 943 nucleotides in length excluding the poly(A) region. An unexpected finding was that there are only three bases in the 5' non-coding region and its significance in terms of ribosome binding is discussed; 87 code for the precursor or leader sequence of the protein, 642 for the mature protein and 211 for the 3' non-coding region. The codons for the precursor region allows the previously undetermined amino acid sequence to be predicted. In common with other precursor regions a high proportion of the predicted amino acids are hydrophobic.

摘要

已使用链终止法和化学合成的脱氧寡核苷酸确定了小鼠免疫球蛋白κ轻链MOPC 21信使RNA的完整序列,以启动与mRNA模板互补的DNA分子的合成。五个这样的寡核苷酸引物已用于该信使RNA的序列分析。该方法对于小鼠κ链mRNA的比较研究非常出色,因为它们可以在不纯的mRNA制剂上制备。MOPC 21轻链mRNA长度为943个核苷酸,不包括聚腺苷酸(poly(A))区域。一个意外的发现是5'非编码区只有三个碱基,并讨论了其在核糖体结合方面的意义;87个编码蛋白质的前体或前导序列,642个编码成熟蛋白质,211个编码3'非编码区。前体区域的密码子使得可以预测先前未确定的氨基酸序列。与其他前体区域一样,预测的氨基酸中有很大一部分是疏水的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6b48/327452/354bd909d1a4/nar00411-0057-a.jpg

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