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人次黄嘌呤磷酸核糖基转移酶全长可表达cDNA的分离与鉴定

Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyl transferase.

作者信息

Jolly D J, Okayama H, Berg P, Esty A C, Filpula D, Bohlen P, Johnson G G, Shively J E, Hunkapillar T, Friedmann T

出版信息

Proc Natl Acad Sci U S A. 1983 Jan;80(2):477-81. doi: 10.1073/pnas.80.2.477.

Abstract

We have cloned a full-length 1.6-kilobase cDNA of a human mRNA coding for hypoxanthine phosphoribosyltransferase (HPRT; IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) into a simian virus 40-based expression vector and have determined its full nucleotide sequence. The inferred amino acid sequence agrees with a partial amino acid sequence determined for authentic human HPRT protein. Transfection of HPRT-deficient mouse LA9 cells with the purified plasmid leads to the expression of human HPRT enzyme activity in cells stably transfected and selected for enzyme activity in hypoxanthine/aminopterin/thymidine medium.

摘要

我们已将编码次黄嘌呤磷酸核糖转移酶(HPRT;肌苷酸:焦磷酸磷酸核糖转移酶,EC 2.4.2.8)的人类mRNA的全长1.6千碱基cDNA克隆到基于猿猴病毒40的表达载体中,并确定了其完整的核苷酸序列。推导的氨基酸序列与为真正的人类HPRT蛋白确定的部分氨基酸序列一致。用纯化的质粒转染缺乏HPRT的小鼠LA9细胞,可导致在次黄嘌呤/氨基蝶呤/胸腺嘧啶核苷培养基中稳定转染并选择具有酶活性的细胞中表达人类HPRT酶活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b241/393401/a79ef9e35195/pnas00628-0165-a.jpg

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