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Development and analysis of secretory component disrupted mouse

分泌成分破坏小鼠的开发和分析

基本信息

批准号：
09357016
负责人：
MORO Itaru
金额：
$ 12.16万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09357016/
关键词：
secretory component mucosal immunity gene mRNA genomic DNA gene targeting polymeric IgA S-IgA Northern blot analysis Secretory Component Disrupted mice ES cell TT2 neomycine PCR法 Southern blotting法キメラマウス

项目摘要

Secretory component (SC), often called polymeric immunoglobulin receptor (pIgR), is a component of SIgA which plays an important role in the mucosal defense system. SC which is localized on the basolateral membrane of the epithelial cells at the mucosal site is a receptor for polymeric immunoglobulins. Accordingly, it transports polymeric immunoglobulin and immune complex through epithelial cells into the secretion. SC is a transmembrane protein with an Mr of 80 kDa abd cDNA revealed 1 lexons composed of extracellular region, transmembrane region and cytoplasmic tail.The purpose of this project is to develop SC disrupted mouse to examine the function of SC.The results obtained were as follows;1) genomic DNA for SC was screened using a mouse pIgR cDNA and was determined by sequencing of positive clones.2) Five positive clones were obtained. The results of restriction enzyme map and Southern blotting revealed a 9.4 kb of these clones. Sequencing analysis indicated that the BamH I-EcoR I fragment was 8404bp, and exon 4, 5, 6, 7, 8, 9, 10 were 653 bp, 352 bp, 332 bp, 192 bp, 129 bp, 131 bp and 58 bp, respectively. Introns 4, 5, 6, 7, 8, 9, 10 were 1237 bp, 552 bp, 891 bp, 359 bp, 368 bp, 1106 bp and 1010 bp, respectively. The size of the exon 11 to EcoR I fragment was 2543 bp.

分泌成分（SC），通常称为多聚免疫球蛋白受体（pIgR），是SIgA的一种成分，在粘膜防御系统中发挥重要作用。位于粘膜部位上皮细胞基底外侧膜上的SC是聚合免疫球蛋白的受体。因此，它将多聚免疫球蛋白和免疫复合物通过上皮细胞转运到分泌物中。SC是一种跨膜蛋白，分子量约为80 kDa，cDNA显示由胞外区、跨膜区和胞质尾区组成的1个单元，本课题旨在建立SC破坏小鼠模型，研究SC的功能。1）使用小鼠pIgR cDNA筛选SC的基因组DNA，并通过阳性克隆的测序确定。2）获得5个阳性克隆。酶切图谱和Southern杂交结果表明，克隆长度为9.4kb。测序结果表明，BamH Ⅰ-EcoR Ⅰ片段长度为8404 bp，外显子4、5、6、7、8、9、10分别为653 bp、352 bp、332 bp、192 bp、129 bp、131 bp和58 bp。内含子4、5、6、7、8、9、10分别为1237 bp、552 bp、891 bp、359 bp、368 bp、1106 bp和1010 bp。外显子11至EcoR Ⅰ片段大小为2543 bp。