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p****c
发帖数: 233 | 1 各位牛人，想请教一个问题如下：
我make了几个mammalian expressing constructs containing FLAG tagged Glycine
domain （150 amino acids ）。The sequencing results are correct。They all
have
KOZAK and are inframe. 但是，当我overexpress them individually in HEKT cells
，
Western blotting can not detect FLAG。 但其他 constructs containing these
Glycine domain fused with another protein domain can be detected by Western
blotting （FLAG）。
我百思不得其解。不知是因为these Glycine domain （150 aa）太小，再HEKT cell
里不稳定
被降解了吗？奇怪的是大蛋白（含有相同的Glycine domain but fu | s******y
发帖数: 28562 | 2 If the sequence contains nothing but glycine, then it is possible it will be
degraded after synthesis or get secreted because it is too hydrophobic
and will tend to aggregate.
cells
Western
【在 p****c 的大作中提到】
: 各位牛人，想请教一个问题如下：
: 我make了几个mammalian expressing constructs containing FLAG tagged Glycine
: domain （150 amino acids ）。The sequencing results are correct。They all
: have
: KOZAK and are inframe. 但是，当我overexpress them individually in HEKT cells
: ，
: Western blotting can not detect FLAG。 但其他 constructs containing these
: Glycine domain fused with another protein domain can be detected by Western
: blotting （FLAG）。
: 我百思不得其解。不知是因为these Glycine domain （150 aa）太小，再HEKT cell
| p****c
发帖数: 233 | 3 Thank you very much, sunnyday. It sounds a very reasonable explanation. I
checked and found there are around 50% of Glycines in these Glycine domains. |
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