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丫头7976

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[求助] 关于在表达蛋白和GFP蛋白之间的linker

各位好,我最近在做膜蛋白的表达,在构建载体的时候我在膜蛋白的C端还加了GFP指示蛋白,但是在诱导表达后,根本看不到菌发出绿色荧光,而我只表达GFP时却能看到菌是绿色的,我在构建载体的时把膜蛋白和GFP直接连在一起了,中间没有加linker regions,我想问一下各位,我的菌株看不出绿色的原因是不是我没有加一个linker的原因呢,如果有做过这方面的大侠是不是可以知道一下我呢,多谢各位!
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丫头7976

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引用回帖:
Originally posted by bio-research at 2011-06-06 11:13:02:
protein of interest surely weakens fluorescence of GFP with linker or without linker.
i have ever made experiment about expression of fusion protein including GFP,
the fusion protein including GF ...

Thank you very much for your advice, but there is a question, when you express fusion protein including GFP,  can you watch the green fluorescence with the naked eye ?
3楼2011-06-06 17:47:22
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丫头7976

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引用回帖:
Originally posted by 西瓜 at 2011-06-06 22:35:00:
You can't see the fluorescence if the GFP is not excitated. But, if your fusion protein is well expressed, you can see that the bacteria expressing the fusion protein is green, without excita ...

thank you for your help, I'd like to make a quest if I may,could you give me  a QQ number? I want to make a more specific communications. I hope you will meet.
6楼2011-06-07 16:18:04
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丫头7976

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dhd997(金币+1): 欢迎来本版继续交流 2011-07-17 09:42:46
引用回帖:
Originally posted by 丫头7976 at 2011-06-07 16:18:04:
thank you for your help, I'd like to make a quest if I may,could you give me  a QQ number? I want to make a more specific communications. I hope you will meet.

你好,我最近又在做膜蛋白与GFP的融合蛋白,我诱导表达后,用肉眼看不出菌体是绿色的,是我的蛋白的表达量少看不出绿色呢,还是说得用激发光激发才能看出绿色呢,你有做过膜蛋白与GFP融合蛋白的表达吗,确定能用肉眼看出菌体是绿色的吗,请用中文回复我吧,英文交流的不是很清楚,谢谢你啦
8楼2011-07-16 21:13:56
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