中文摘要：

[目的]构建鼠Ⅰ型胶原蛋白α1链（COL1A1）C端甘氨酸重复序列（氨基酸1 000~1 189）的真核表达载体,探究其与成纤维细胞（Rat-1）的相互作用。[方法]以COL1A1 c DNA为模板,PCR扩增甘氨酸重复序列基因,克隆至p AP-tag4载体上得到重组质粒,命名为p AP-Gly。将p AP-Gly转染至CHO细胞,Western Blot鉴定目的蛋白。用细胞原位和定量染色及配体受体亲和印迹实验探究其与Rat-1细胞的相互作用。[结果]p AP-Gly成功构建并表达,Western Blot检测到77k Da的目的条带。AP-Gly与Rat-1的原位染色及定量染色相对OD405值（OD405=1.157±0.066）表明其能与Rat-1结合。[结论]p AP-Gly成功构建并表达,融合蛋白能和Rat-1表面相互作用,为研究Ⅰ型胶原蛋白在成纤维细胞表面上的受体复合物奠定了基础。

英文摘要：

[ Objective]To construct the eukaryotic expression vector of C -terminal glycine repeats （amino acids 1 000 - 1 189） of rat collagen type 1 alpha 1 （COL1A1） and observe the interaction between the expressed product and fibroblasts （Rat- 1 ）. [ Methods ] The glyeine repeats gene from the COL1 A1 cDNA sequence was amplified by PCR and cloned into ex- pression vector pAP - tag4, and the recombinant was designated pAP - Gly. The right recombinant plasmid was transfected into CHO cells. Western Blot was performed for further confirmation. In situ and quantitative binding assays as well as ligand - re- ceptor affinity experiments were performed to investigate the interplay between the fusion protein and Rat - 1 cells. [ Results ] The plasmid pAP - Gly was successfully constructed and expressed and western blot analysis showed the 77 kDa target protein. In situ and quantitative binding assay （relative OD4o5 = 1. 157 ＋0.066） demonstrated that AP- Gly could bind to the surface of Rat- 1 cells. [ Conclusion]The recombinant pAP- Gly was successfully constructed and expressed,and the fusion protein could interplay with Rat - 1 cells which provides a foundation for further exploring the receptor complexes of COL1A1 on the surface of fibroblasts.