Chardon P, Kirszenbaum M, Cullen P R, Geffrotin C, Auffray C, Strominger J L, Cohen D, Vaiman M
Immunogenetics. 1985;22(4):349-58. doi: 10.1007/BF00430918.
Four cDNA probes for the human major histocompatibility complex (MHC) were used to investigate the sheep MHC, in conjunction with serological typing for ovine lymphocyte antigen (OLA). Lymphocytes from a family (two parents and five offspring) of Romanov sheep were subjected to genomic DNA digestion by the restriction endonuclease Eco RI, followed by gel electrophoresis. A single Southern blot representing all seven individuals was then consecutively hybridized with the class I, alpha-DC, beta-DR, and C4 probes, which were originally designed to identify HLA class I, class II (DC and DR), and C4 products, respectively. Using each of the three class I/class II probes, several bands showing DNA polymorphism were detected. The segregation of these bands in the five offspring exactly paralleled the OLA haplotype segregation established by serological typing. A further eight individuals carrying haplotypes which were phenotypically identical to those in the above-mentioned family showed bands in the corresponding positions when tested with the same three probes. Using the C4 probe, no polymorphism was detected in these fifteen individuals.
使用四种针对人类主要组织相容性复合体(MHC)的cDNA探针,结合绵羊淋巴细胞抗原(OLA)的血清学分型,对绵羊MHC进行研究。来自罗曼诺夫羊一个家系(两只亲本和五只后代)的淋巴细胞用限制性内切酶Eco RI进行基因组DNA消化,然后进行凝胶电泳。接着,将代表所有七个个体的单一Southern印迹依次与I类、α-DC、β-DR和C4探针杂交,这些探针最初分别设计用于鉴定HLA I类、II类(DC和DR)以及C4产物。使用三种I类/II类探针中的每一种,都检测到了几条显示DNA多态性的条带。这五条带在五个后代中的分离情况与通过血清学分型确定的OLA单倍型分离情况完全平行。另外八个携带与上述家系表型相同单倍型的个体,在用相同的三种探针进行检测时,在相应位置出现了条带。使用C4探针,在这十五个个体中未检测到多态性。