Wu M S, Tani K, Sugiyama H, Hibino H, Izawa K, Tanabe T, Nakazaki Y, Ishii H, Ohashi J, Hohjoh H, Iseki T, Tojo A, Nakamura Y, Tanioka Y, Tokunaga K, Asano S
Division of Molecular Therapy, Advanced Clinical Research Center (ACRC), The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
J Mol Evol. 2000 Sep;51(3):214-22. doi: 10.1007/s002390010083.
A New World monkey, the common marmoset (Callithrix jacchus), will be used as a preclinical animal model to study the feasibility of cell and gene therapy targeting immunological and hematological disorders. For elucidating the immunogenetic background of common marmoset to further studies, in the present study, polymorphisms of MHC-DRB genes in this species were examined. Twenty-one Caja-DRB exon 2 alleles, including seven new ones, were detected by means of subcloning and the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods followed by nucleotide sequencing. Based on the alignment of these allele sequences, we designed two pairs of specific primers and established a PCR-SSCP method for DNA-based histocompatibility typing of the common marmoset. According to the family segregation data and phylogenetic analyses, we presumed that Caja-DRB alleles could be classified into five different loci. Southern blotting analysis also supported the existence of multiple DRB loci. The patterns of nucleotide substitutions suggests that positive selection operates in the antigen-recognition sites of Caja-DRB genes.
普通狨猴（Callithrix jacchus）作为一种新世界猴，将被用作临床前动物模型，以研究针对免疫和血液系统疾病的细胞和基因治疗的可行性。为了阐明普通狨猴的免疫遗传背景以便进一步研究，在本研究中，检测了该物种MHC-DRB基因的多态性。通过亚克隆和聚合酶链反应-单链构象多态性（PCR-SSCP）方法，随后进行核苷酸测序，检测到21个Caja-DRB外显子2等位基因，其中包括7个新的等位基因。基于这些等位基因序列的比对，我们设计了两对特异性引物，并建立了一种用于普通狨猴基于DNA的组织相容性分型的PCR-SSCP方法。根据家系分离数据和系统发育分析，我们推测Caja-DRB等位基因可分为五个不同的位点。Southern印迹分析也支持多个DRB位点的存在。核苷酸替换模式表明，正选择作用于Caja-DRB基因的抗原识别位点。
