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绵羊主要组织相容性复合体II类(OLA-DR)基因的表达与特性分析

Expression and characterization of ovine major histocompatibility complex class II (OLA-DR) genes.

作者信息

Ballingall K T, Wright H, Redmond J, Dutia B M, Hopkins J, Lang J, Deverson E V, Howard J C, Puri N, Haig D

机构信息

Moredun Research Institute, Edinburgh, UK.

出版信息

Anim Genet. 1992;23(4):347-59. doi: 10.1111/j.1365-2052.1992.tb00157.x.

Abstract

Previous work made use of nucleic acid probes corresponding to different subtypes of the class II regions of the human and murine major histocompatibility complex (MHC) to isolate seven different alpha and 24 different beta genes of the ovine MHC from two cosmid libraries. In an attempt to identify pairs of alpha and beta genes capable of cell surface expression, all permutations of alpha and beta genes were in turn transfected into mouse L-cells. Two pairs of alpha and beta genes co-expressed and stable ovine MHC class II L-cell lines were developed. The expressed alpha genes had previously been defined as DR-alpha homologues (DRA) by differential Southern hybridization to human subtype specific class II probes. The expressed ovine beta genes were also assigned as ovine DR-beta homologues (DRB) on the basis of their sequence having a higher degree of similarity with human DRB than any other subtype. A total of eight out of 23 anti-sheep class II specific monoclonal antibodies were typed OLA-DR specific by FACScan analysis using the L-cell lines.

摘要

先前的研究利用与人类和小鼠主要组织相容性复合体(MHC)Ⅱ类区域不同亚型相对应的核酸探针,从两个黏粒文库中分离出绵羊MHC的7种不同的α基因和24种不同的β基因。为了鉴定能够在细胞表面表达的α和β基因对,将α和β基因的所有排列组合依次转染到小鼠L细胞中。构建了两对共同表达且稳定的绵羊MHCⅡ类L细胞系。通过与人类亚型特异性Ⅱ类探针进行差异Southern杂交,先前已将表达的α基因定义为DR-α同源物(DRA)。根据表达的绵羊β基因序列与人类DRB的相似程度高于任何其他亚型,这些β基因也被指定为绵羊DR-β同源物(DRB)。使用这些L细胞系,通过FACScan分析,在23种抗绵羊Ⅱ类特异性单克隆抗体中,共有8种被鉴定为OLA-DR特异性。

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