Access to a messenger RNA sequence or its protein product is not limited by tissue or species specificity.
Access to a messenger RNA sequence or its protein product is not limited by tissue or species specificity.
复制标题
信使 RNA 序列或其蛋白质产物的获取不受组织或物种特异性的限制。
DOI：
10.1126/science.2565599
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发表时间：
1989
期刊：
影响因子：
56.9
通讯作者：
Steve S. Sommer
中科院分区：
文献类型：
--
作者：
Gobinda Sarkar;Steve S. Sommer
RNA扩增与转录序列（RAWTS）是一种快速、灵敏的直接测序方法，涉及互补DNA合成、用含有噬菌体启动子的引物进行聚合酶链反应（PCR）、从噬菌体启动子转录以及逆转录介导的测序。通过RAWTS，可以在检测的四种细胞类型（白细胞、肝脏、K562红白血病细胞和绒毛膜绒毛细胞）中对四种组织特异性人类信使rna（蓝色色素、因子IX、苯丙氨酸羟化酶和酪氨酸羟化酶）进行测序。这些结果表明，在成人和胚胎组织中存在组织特异性mrna的转录、剪接和多聚腺苷化的基本速率。除了揭示序列信息外，还可以通过将翻译起始信号整合到适当的PCR引物中来产生所需的体外翻译产物。RAWTS可用于从其他物种获得新的mRNA序列信息，如因子IX的催化结构域片段所示。总的来说，快速获得跨物种mRNA序列的能力应该有助于蛋白质进化的研究和鉴定对蛋白质结构和功能至关重要的序列。
RNA amplification with transcript sequencing (RAWTS) is a rapid and sensitive method of direct sequencing that involves complementary DNA synthesis, polymerase chain reaction (PCR) with a primer or primers containing a phage promoter, transcription from the phage promoter, and reverse transcriptase-mediated sequencing. By means of RAWTS, it was possible to sequence each of four tissue-specific human messenger RNAs (blue pigment, factor IX, phenylalanine hydroxylase, and tyrosine hydroxylase) in four cell types examined (white blood cells, liver, K562 erythroleukemia cells, and chorionic villus cells). These results indicate that there is a basal rate of transcription, splicing, and polyadenylation of tissue-specific mRNAs in adult and embryonic tissues. In addition to revealing sequence information, it is possible to generate a desired in vitro translation product by incorporating a translation initiation signal into the appropriate PCR primer. RAWTS can be used to obtain novel mRNA sequence information from other species as illustrated with a segment of the catalytic domain of factor IX. In general, the ability to obtain mRNA sequences rapidly across species boundaries should aid both the study of protein evolution and the identification of sequences crucial for protein structure and function.
影响因子：
2.9
作者：
YOSHITAKE, S;SCHACH, BG;KURACHI, K
通讯作者：
KURACHI, K