LTB-TGFβ1融合蛋白原核表达质粒的构建

LTB-TGFβ1融合蛋白原核表达质粒的构建(7700字)
摘  要：转化生长因子( TGFβ1)是一种强烈的免疫抑制因子，近年来还发现其可诱导调节性T细胞的生成而介导特异性免疫耐受，在家畜的持续性感染过程中发挥重要的作用，抑制TGFβ1作用可望增强机体的免疫功能。本实验拟应用基因工程技术制备LTB-TGFβ1融合蛋白，作为TGFβ1自体疫苗，来诱导机体产生抗TGFβ1抗体，探讨抑制TGFβ1是否有利于机体免疫系统清除持续性感染的病原体。方法：采用人工合成LTB-柔性连接-TGFβ1编码区DNA，连接于pUC57 T载体，经双酶切后回收目的片段连接于pET28载体；结果：经双酶切及DNA序列分析证实融合基因序列完全正确；结论:本研究成功构建了LTB-TGFβ1融合基因，这为下一步进行LTB-TGFβ1的原核表达打下了基础。
关键词：转化生长因子β1 (TGF-β1)；大肠杆菌不耐热肠毒素B；融合蛋白；原核表达
                                       
LTB - TGFβ1Fusion Protein Expression Vector
Abstract: The transforming growth factor (TGFβ1) is a strong immunosuppressive factor in recent years also found that regulatory T cells can induce the formation of the induced specific immune tolerance, persistent infection in domestic animals play an important process role, the role of inhibition of TGFβ1 is expected to enhance immune function. This study was to genetic engineering preparation LTB-TGFβ1 fusion protein, as TGFβ1 autologous vaccine to induce antibodies against TGFβ1, TGFβ1 inhibition of the immune system is beneficial to clear the persistent infection of pathogens. Methods: The synthetic LTB-flexible connection-TGFβ1 encoding DNA, connected to pUC57 T vector, double digested fragment was recovered connected pET28 vector. Results: double digestion and DNA sequence analysis confirmed the correct fusion gene sequences. Conclusion: The successfully constructed LTB-TGFβ1 fusion gene, which is the next step for LTB-TGFβ1 expression original foundation.
Key words: transforming growth factor β1 (TGF-β1); Escherichia coli heat-labile enterotoxin B; fusion protein; prokaryotic expression