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树鼩作为多种人类疾病研究模型的可能性已受到广泛关注,但尚缺乏研究其免疫功能的基本标志以及单克隆抗体。该实验首先以树鼩外周血总RNA为材料,通过RT-PCR扩增得到长度为1365bp的树鼩CD4全长编码序列,并确定了数据库中缺失的两个片段,进而通过ClustalW等软件对其序列和分子特征进行分析,发现树鼩CD4氨基酸序列胞外和胞内域保守性较好,且与人类和猴的亲缘关系较近。虽然树鼩和人CD4分子表面大部分区域均带正电荷,但与人CD4胞外域D1相比,树鼩CD4D1结构区域表面带负电荷较多,且多出两个N-糖基化位点。这些差异对抗体的结合可能存在影响。该研究为今后树鼩CD4单克隆抗体制备及功能研究奠定了基础。
The possibility of tree shrew as a model for a variety of human diseases has received widespread attention, but there is a lack of basic markers for studying its immune function as well as monoclonal antibodies. In this experiment, we first obtained the full-length coding sequence of 1365bp in tree shrews from total RNA of tree shrews by RT-PCR and identified the missing two fragments in the database. Sequence and molecular characteristics of the tree shrews CD4 amino acid sequence found in the extracellular and intracellular domains of conservation is better, and with human and monkey closer genetic relationship. Although the majority of the surface area of the CD4 子 and human CD4 molecules are positively charged, compared with the human CD4 extracellular domain D1, the surface structure of the CD4D1 structural region of the tree 带 is more negatively charged and more two N-glycosylation sites . These differences may affect the binding of antibodies. This study lays the foundation for the preparation and function study of CD4 monoclonal antibody in tree shrews in the future.