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利用单克隆抗体克隆编码与光系统I相关的捕光复合体多肽的豌豆cDNA。

Cloning of a pea cDNA encoding a polypeptide of the light-harvesting complex associated with photosystem I using a monoclonal antibody.

作者信息

Cannell M E, Mitchell A J, McCready S, Callow J A, Green J R

机构信息

School of Biological Sciences, University of Birmingham, UK.

出版信息

Plant Mol Biol. 1995 Feb;27(4):821-4. doi: 10.1007/BF00020235.

Abstract

A monoclonal antibody (MAb UB42) is described that binds to thylakoids in pea chloroplasts, as shown by EM-immunogold labelling. The antibody recognised proteins of ca. 23-29 kDa in western blots of a pea leaf homogenate. A cDNA library was prepared from pea epidermal cells in the vector lambda ZAP II, and immunoscreening of the library with UB42 led to the isolation of a clone, pUB42. This was sequenced and had an open reading frame of 269 codons encoding a predicted polypeptide of 28.9 kDa. The sequence showed extensive homology with three closely related polypeptides belonging to a family of chlorophyll a/b-binding proteins from the light harvesting complex of photosystem I (LHCI). Collectively, the results suggest that MAb UB42 recognises an epitope on the type II chlorophyll a/b-binding protein from LHCI and that clone pUB42 encodes this protein.

摘要

描述了一种单克隆抗体(MAb UB42),如免疫电镜金标所示,它能与豌豆叶绿体类囊体结合。在豌豆叶片匀浆的蛋白质印迹法中,该抗体识别约23 - 29 kDa的蛋白质。用载体λZAP II从豌豆表皮细胞制备了一个cDNA文库,用UB42对该文库进行免疫筛选,得到一个克隆pUB42。对其进行测序,有一个269个密码子的开放阅读框,编码一个预测分子量为28.9 kDa的多肽。该序列与来自光系统I捕光复合体(LHCI)的叶绿素a/b结合蛋白家族的三个密切相关的多肽具有广泛同源性。总体而言,结果表明MAb UB42识别LHCI中II型叶绿素a/b结合蛋白上的一个表位,且克隆pUB42编码该蛋白。

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