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人纤溶酶原全长cDNA克隆的分子克隆与鉴定

Molecular cloning and characterization of a full-length cDNA clone for human plasminogen.

作者信息

Forsgren M, Råden B, Israelsson M, Larsson K, Hedén L O

出版信息

FEBS Lett. 1987 Mar 23;213(2):254-60. doi: 10.1016/0014-5793(87)81501-6.

Abstract

A human liver cDNA library enriched for full-length clones was screened for plasminogen cDNA using a synthetic 24-nucleotide probe derived from a reported partial cDNA sequence. 12 positive clones were identified and one of these was characterized in detail. The 2.7 kb insert contains the complete coding region. At 5 positions, it gives residues different from those reported in a previous amino acid sequence analysis of the protein. The present results show an extra Ile at position 65, Gln instead of Glu at positions 53 and 342, Asn at position 88 instead of Asp, and Asp at position 453 rather than Asn. In the 3'-non-coding region an extension of 29 bases is found which does not contain any structure compatible with a known polyadenylation signal. Instead, the consensus signal AATAAA is placed at a distance of 46 bases upstream of the poly(A)-tail.

摘要

利用从已报道的部分cDNA序列推导而来的24个核苷酸的合成探针,对富含全长克隆的人肝脏cDNA文库进行纤溶酶原cDNA筛选。鉴定出12个阳性克隆,并对其中一个进行了详细表征。2.7kb的插入片段包含完整的编码区。在5个位置上,其给出的残基与先前该蛋白质的氨基酸序列分析报道的不同。目前的结果显示,第65位有一个额外的异亮氨酸,第53位和342位为谷氨酰胺而非谷氨酸,第88位为天冬酰胺而非天冬氨酸,第453位为天冬氨酸而非天冬酰胺。在3'-非编码区发现了一个29个碱基的延伸,其中不包含任何与已知聚腺苷酸化信号相容的结构。相反,共有信号AATAAA位于聚(A)尾上游46个碱基处。

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