Au D M, Wong W M, Tam J W, Cheng L Y, Lam V M
Department of Biochemistry, University of Hong Kong, Hong Kong.
Gene. 1995 Nov 20;165(2):261-6. doi: 10.1016/0378-1119(95)00570-v.
The isolation and detailed characterization of a three-beta-globin gene (GloB) haplotype in the Sprague-Dawley (S-D) rat is described. An enriched library, lambda SDHelib, was screened with a human GloB probe, humbg44, and from which a beta minor gene, Rathbbz, was isolated, sequenced and characterized. A S-D rat GloB-specific probe, Ratbgze12, derived from the Rathbbz gene, was then used to screen a S-D rat genomic library, lambda SDglib. The clone T1510 was isolated and identified to include the entire Rathbbz gene and part of another GloB gene, Rathbby, which was 5' upstream from Rathbbz. Chromosomal walking upstream using the riboprobe, rnaT71, led to the isolation of an overlapping clone, Ta49, which was shown to include two full-length GloB genes; the most 5' was Rathbbx followed by Rathbby. Sequence data suggests that Rathbbx is a beta major gene, whereas Rathbby is a hybrid gene of Rathbbx and Rathbbz. Genomic hybridization confirmed this particular three-gene haplotype in the S-D rat. This haplotype, a1, may be the prototype of the GloB cluster in rat.
本文描述了在斯普拉格-道利（S-D）大鼠中一个三β-珠蛋白基因（GloB）单倍型的分离及详细特征分析。用人类GloB探针humbg44筛选了一个富集文库λ SDHelib，并从中分离出一个次要β基因Rathbbz，进行了测序和特征分析。然后，用源自Rathbbz基因的S-D大鼠GloB特异性探针Ratbgze12筛选S-D大鼠基因组文库λ SDglib。分离并鉴定出克隆T1510，它包含整个Rathbbz基因和另一个GloB基因Rathbby的一部分，Rathbby位于Rathbbz的5'上游。使用核糖探针rnaT71向上游进行染色体步移，导致分离出一个重叠克隆Ta49，它包含两个全长GloB基因；最5'端的是Rathbbx，接着是Rathbby。序列数据表明，Rathbbx是一个主要β基因，而Rathbby是Rathbbx和Rathbbz的杂交基因。基因组杂交证实了S-D大鼠中这种特定的三基因单倍型。这种单倍型a1可能是大鼠GloB簇的原型。
