人血小板mRNA的分离、鉴定及λgt11 cDNA文库的构建。通过鉴定糖蛋白Ib（GPIb）编码mRNA和克隆GPIb编码cDNA插入片段来确认血小板来源。
Isolation and characterization of human blood platelet mRNA and construction of a cDNA library in lambda gt11. Confirmation of the platelet derivation by identification of GPIb coding mRNA and cloning of a GPIb coding cDNA insert.
作者信息
Wicki A N, Walz A, Gerber-Huber S N, Wenger R H, Vornhagen R, Clemetson K J
机构信息
Theodor Kocher Institute, Bern, Switzerland.
出版信息
Thromb Haemost. 1989 Jun 30;61(3):448-53.
We have developed a purification method for the isolation of platelet-specific poly (A+) RNA and demonstrated that human blood platelets, despite the absence of a nucleus, contain stable mRNA. The poly (A+) RNA was used to construct a platelet-specific cDNA expression library in lambda gt11. The platelet derivation of the purified mRNA was confirmed by identification of membrane glycoprotein Ib (GPIb) message by immunoprecipitation of rabbit reticulocyte lysate translation products with poly- and monoclonal antibodies against GPIb alpha and by sequencing of a GPIb alpha cDNA clone.
我们已经开发出一种用于分离血小板特异性聚腺苷酸（poly(A+)）RNA的纯化方法，并证明人血血小板尽管没有细胞核，但仍含有稳定的信使核糖核酸（mRNA）。利用该聚腺苷酸（poly(A+)）RNA在λgt11载体中构建了血小板特异性互补脱氧核糖核酸（cDNA）表达文库。通过用抗糖蛋白Ibα（GPIbα）的多克隆和单克隆抗体对兔网织红细胞裂解物翻译产物进行免疫沉淀来鉴定膜糖蛋白Ib（GPIb）信息，并对GPIbα互补脱氧核糖核酸（cDNA）克隆进行测序，从而证实了纯化信使核糖核酸（mRNA）的血小板来源。
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