大家帮忙看看这个GFP融合蛋白怎么了？ - 未名空间MITBBS历史存档

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大家帮忙看看这个GFP融合蛋白怎么了？

大家帮忙看看这个GFP融合蛋白怎么了？# Biology - 生物学

s*m2012-02-21 08:02

1 楼

谢谢！

z*i2012-02-21 08:02

2 楼

有人试过在Alexandria, VA walk in么？多谢！

f*g2012-02-21 08:02

3 楼

如题谢谢

H*32012-02-21 08:02

4 楼

我的情况跟前一段时间讨论的一个情况不同。
转基因植物携带overexpression promoter+gene+c terminal GFP，转到野生型里建立
过量表达突变体。基因是核编码的叶绿体蛋白，蛋白N末端一个小酞段引导蛋白
targeting到叶绿体内行使功能。根据抗性筛选到homozygotes,confocal下检测到GFP信
号在叶绿体里，非常阳性。DNA genotyping和RT-PCR检测到基因插入正常，mRNA正常，
现在的问题是western用anti－GFP抗体无法检测到全长蛋白，只有GFP大概27kD左右的
信号。因为是未知蛋白，没有特异性抗体，突变体性状也没有发表过，所以无法确定是
否融合蛋白正确表达。我唯一能想到的是蛋白合成后，送到叶绿体内被降解了。可是即
使有降解，不会全都降解吧。
请大家给点意见，好不容易建立的突变体，实在不想放弃。

C*n2012-02-21 08:02

5 楼

问一下，你知道discover的网站吗？
谢谢！

h*y2012-02-21 08:02

6 楼

Yes, you can. Just bring your finger print notice and ID with you.

y*n2012-02-21 08:02

7 楼

降解 is very possible.

s*l2012-02-21 08:02

8 楼

http://www.discovercard.com/customer-service/rewards/shopdiscov

z*i2012-02-21 08:02

9 楼

Is it busy there? It took me over 70 days to receive the FP notice!

【在 h***y 的大作中提到】

: Yes, you can. Just bring your finger print notice and ID with you.

H*g2012-02-21 08:02

10 楼

How big is the linker region between your gene of interest and the GFP?

h*y2012-02-21 08:02

11 楼

I went before the new year in the middle in the middle of the day and there
were hardly anybody there. It took me less than 10 mins to get it done. Try
to go in the middle of the day.

H*32012-02-21 08:02

12 楼

20 amino acids. Is it important?

z*i2012-02-21 08:02

13 楼

Thank you so much for your information!

there
Try

【在 h***y 的大作中提到】

: I went before the new year in the middle in the middle of the day and there
: were hardly anybody there. It took me less than 10 mins to get it done. Try
: to go in the middle of the day.