论文部分内容阅读
目的 :初步分析小鼠α2 ( )胶原基因上游 5′- U TR近端 80 0 bp(- 780～ + 5 4bp)序列的 DNA结合蛋白。 方法 :PCR扩增小鼠α2 ( )胶原基因 - 2 5 8～ + 5 4bp,- 5 19～ - 2 48bp,- 741～ - 5 0 1bp片段 ,Dig- d U TP末端标记 PCR产物 ,与经 SDS- PAGE并转印至膜上的胶原产生细胞的核抽提物进行 DNA -蛋白质结合反应 ,以抗 Dig抗体检测结合反应信号。 结果 : 型胶原基因上游 5′- U TR近端 80 0 bp序列中存在至少 3个不同的核转录因子结合位点 ,相识别 (consensus)的核蛋白因子相对分子质量分别为 12万 ,5万 ,2万。TGF- β1可增强细胞中相对分子质量为 12万的核蛋白因子的表达。结论 :小鼠 α2( )胶原基因上游 - 780～ + 5 4bp这 80 0 bp片段中存在与不同的核蛋白因子结合的序列 ,可能与 Sp- 1有关。TGF- β1通过增强相对分子质量为 12万核蛋白的表达而促进该蛋白与靶调控序列的结合。
OBJECTIVE: To analyze the DNA binding protein sequence of 80 0 bp (-780 ~ + 5 4bp) proximal to 5’-U TR upstream of mouse α2 () collagen gene. Methods: The PCR products of mouse α2 (-) collagen gene-2 5 8 ~ + 5 4 bp, -5 19 ~ -2 48 bp, - 741 ~ -5 0 1 bp fragment were amplified by PCR. SDS-PAGE and transferred to membrane-derived collagen-producing cell nuclear extracts for DNA-protein binding reactions, with anti-Dig antibody binding reaction signal. Results: There were at least 3 different nuclear transcription factor binding sites in the 80 bp sequence of the 5’-U TR upstream of the type collagen gene. The relative molecular mass of the consensus nucleoprotein factors were 120,000 and 50,000 respectively ,20000. TGF-β1 enhances the expression of nucleoprotein factors with a relative molecular mass of 120,000 in cells. CONCLUSION: The upstream of the α2 (superscript +) collagen gene in mouse - 780 ~ + 5 4bp fragment has a binding sequence with different nucleoprotein factors, which may be related to Sp-1. TGF-β1 promotes the binding of the protein to target regulatory sequences by enhancing the expression of a molecular weight of 120,000 nucleoprotein.