【摘 要】
：
目的:完成钝尾毒蜥Exendin -4基因在大肠杆菌中的串联高效表达.方法:按照大肠杆菌偏爱密码子设计Exendin -4基因片段,利用同尾酶构建多拷贝串联的表达载体,于大肠杆菌BL21( D
【机 构】
：
山东省内分泌与代谢病研究所山东省医学科学院
论文部分内容阅读
目的:完成钝尾毒蜥Exendin -4基因在大肠杆菌中的串联高效表达.方法:按照大肠杆菌偏爱密码子设计Exendin -4基因片段,利用同尾酶构建多拷贝串联的表达载体,于大肠杆菌BL21( DE3)中诱导表达,SDS - PAGE鉴定结果,表达产物Ni柱纯化后肠激酶切割,高效液相色谱及四级杆-飞行时间质谱纯化、鉴定,确定目标产物,作用胰岛瘤细胞INS -1检测胰岛素释放活性.结果:成功构建重组载体pET28a -(Exendin -4)n(n=2,4,6),且均获得可溶性表达产物,重组表达蛋白经切割、纯化、鉴定和制备,最终得到纯度98%的Exendin -4,其具有与标准品相似的促葡萄糖刺激的胰岛素释放活性.结论:试验成功利用串联表达方式制备有活性的Exendin -4,为下一步2型糖尿病治疗药物的研究奠定了基础.
其他文献
Anammox was successfully transited from heterotrophic denitrification and autotrophic denitrification in two upflow biofilm reactors, respectively. The results
质粒DNA转化细菌是分子生物学中最常用的技术.经典的氯化钙转化法操作复杂且效率低;通过病毒转化质粒虽然效率高但对健康有威胁;电穿孔基因转移法是目前最有前途的一种方法,
A Particle Image Velocimetry (PIV) method based on the image separation and reconstruction with the median filter and triangular Bézier patch was proposed to m
Effects of bioremediation on microbial communities in soils contaminated with petroleum hydrocarbons are a scientific problem to be solved. Changes in dominate
Simultaneous biological phosphorus and nitrogen removal with enhanced anoxic phosphate uptake via nitrite was investigated in an anaerobic-aerobic-anoxic-aerobi
Forced oxidation of calcium sulfite aqueous slurry is a key step for the calcium-based flue gas desulfurization(FGD) residue. Experiments were conducted in a se
Crimean-Congo Haemorrhagic Fever Virus(CCHFV)is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes,high fat
The basic pyrolysis behaviour of ethylene vinyl acetate(EVA) copolymer, which is often used as a lamination agent in solar modules, was investigated in thermogr
In order to improve nitrogen removal in anoxic/oxic(A/O) process effectively for treating domestic wastewaters, the influence factors, DO(dissolved oxygen), nit
Behaviors of soil heavy metals are often affected by coexisting herbicides due to their physical and chemical interaction. Effect of glyphosate, an herbicide co