基因工程第三章 分子克隆载体.ppt

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * 1 2 3 4 Fig2.1 SDSanalysis of soluble CBD-LTΔ27 after induction 1.Protein molecular weight marker 2.Total bacterial protein, uninduced 3.Total bacterial protein, 30℃ induced 4.Soluble fraction, 30℃ induced 摇瓶发酵生产可溶性融合蛋白的SDS分析 1 2 3 4 5 6 7 8 Fig2.2 SDSanalysis of CBIND 100 Resin purified soluble fusion protein 1.Protein molecular weight marker 5.Supernatant after wash buffer washing 2.Soluble bacterial protein, 30℃ induced 6.Supernatant of first eluting 3.Supernatant after CBD resin binding 7.Supernatant of second eluting 4 Supernatant after bind buffer washing 8.Supernatant of third eluting CBIND 100 Resin纯化可溶表达的融合蛋白 1 2 3 4 5 6 Fig2.3 SDSanalysis of fusion protein lysed by EK 1.Protein molecular weight marker 2.Fusion protein, CBD resin purified 3.EK, 12hr 4.EK, 16hr 5.EK, 20hr 6.EK, 24hr 融合蛋白经肠激酶酶切不同时间的SDS分析 1 2 3 4 1 2 3 4 Fig2.4 SDSand Western blot of purified product LTΔ27 1. Marker 2. Fusion protein, CBD resin purified 3. EK, 16hr 4. LTΔ27, Ni2+ resin purified Fig 2.4a Fig 2.4b 纯化产物的SDS及Western 印迹分析 N- and C-terminal Intein Fusions N- -terminal Intein Fusions Fusion of the intein-tag to the N-terminus of the target protein and release of the target by pH and temperature induced cleavage. PerR protein t