肝脏特异性IgD高表达转基因小鼠的构建  

Establishment of Transgenic Mouse Model with High-level Liver-specific Expression of IgD

作　　者：张希瑶 王喜凤[2] 潘庆杰[1] 闵令江[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]中国农业大学生物学院,北京100093

出　　处：《青岛农业大学学报（自然科学版）》2015年第3期157-162,共6页Journal of Qingdao Agricultural University(Natural Science)

摘　　要：HIDS(高IgD综合征)的引发可能与IgD的升高有关。本试验利用5’RACE技术和免疫球蛋白可变区基因的特异性扩增方法来构建小鼠sIgD(分泌型IgD)重链与轻链可变区基因,利用3’RACE扩增得到sIgD重链与轻链的恒定区基因,融合PCR方法得到sIgD全长。我们分别构建了eAFPpALB-sIgDH、eAFPpALB-IgL、eAFPpALB-GFP和eApoEpAAT-sIgDH、eApoEpAAT-IgL、eApoEpAAT-GFP载体,先进行细胞水平上的初步检测,接着构建了转基因小鼠。对转基因小鼠的检测得知,sIgD在肝脏和脾脏中RNA水平上有少量表达,后续对转基因小鼠各个脏器的RCR及Q-PCR检测发现,sIgD在肾脏中有的较高表达。Hyperimmunoglobulinemia D syndrome（HIDS）may be associated with high level of IgD.Our study used the 5＇RACE technique and specific amplification of rearranged/expressed immunoglobulin variable region genes to build the variable region of heavy and light chain of mouse IgD.Meanwhile,we used3＇RACE technique to build the constant region of heavy and light chain.Then we obtained complete genes of IgD through fusion of PCR.We built six different vectors,such as eAFPpALB-sIgDH、eAFPpALBIgL、eAFPpALB-GFP and eApoEpAAT-sIgDH、eApoEpAAT-IgL and eApoEpAAT-GFP,and did preliminary investigation at cellular level.With those vectors,we generated transgenic mice successfully.Further detection of genetically modified mice showed that there were low levels of expression of sIgD in liver and spleen at RNA level.The follow-up PCR and Q-PCR for viscera of transgenic mice indicated that high levels of expression of sIgD could also be detected in kidney.

关 键 词：IGD 高IgD综合征(HIDS) 转基因小鼠 

分 类 号：Q786[生物学—分子生物学]