j*****y 发帖数: 94 | 1 我现在的蛋白是个his-tagged homo dimer,我想把 WT 和 active-site mutant 放在
一起组成一个heterodimer.
请问大虾有没有什么好的办法能够得到这样一个homogeneous population 的
heterodimer ?
谢谢 |
s********n 发帖数: 2939 | 2 理论上可以这样:
构建两个带有不同purification tags而且可以相容的载体,如6xHis tag on pET和
Strep tag on p15A or dual vector。让这两个monomer coexpress,用Ni-NTA和Strep
tag的亲和柱顺序纯化即可。 |
j*****y 发帖数: 94 | 3 谢谢楼上的。
我的这个蛋白WT和active-site mutant都是homo-dimer,这样的两个载体co-express能
得到heterodimer吗?会不会只能得两种homodimer?还是两种homodimer加上
heterodimer(minor species)? |
s********n 发帖数: 2939 | 4 我说的这个方法正是可以解决你的问题,你仔细想一想。 |
j*****y 发帖数: 94 | |
a**y 发帖数: 464 | 6 you can also use a linker sequence with suitable length to connect one copy
of each gene. |
s******y 发帖数: 28562 | 7 That is what I would prefer. Some dimer proteins (such as the TdtomatoFP)
was made by this strategy
But smartkevin's method is more simple and straightforward
copy
【在 a**y 的大作中提到】![](/moin_static193/solenoid/img/up.png) : you can also use a linker sequence with suitable length to connect one copy : of each gene.
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