C***2 发帖数: 62 | 1 Hey, I am going to generate GeneA+GeneB Knockin mouse in one locus.Furin
cleavage site plus P2A or T2A linker are chosen to link GeneA and GeneB.
This way, co-translation of GenaA and GeneB under endogenous promoter should
be achieved. I googled Furin cleavage site sequence, but i can't find out
accurate and reliable sequence ever used in vivo. So, if someone used or
know furin cleavage site, would you please send me the information and
reference? Moreover, i just read the furin cleavage, do... 阅读全帖 |
|
G***G 发帖数: 16778 | 2 hi
can anyone give us a definition of co-expression?
suppose we have two gene expression across 6 samples.
geneA expression={a,b,c,d,e,f}
geneB expression={A,B,C,D,E,F}
if corr(geneA,geneB) = 1, we say they are co-expressed.
if it is -1, can we still call it as co-expressed? |
|
m******5 发帖数: 1383 | 3 在一个endogenous site
想knock-in一个 GeneA-IRES-GeneB-polyA
理想状况下Gene A和GeneB会在一个大mRNA上分别translate出来
现在面临的问题是,理论上说,核内有一套和核外不同的non sense mediated decay机
制,负责降解stop code和polyA离得太远的情况。 在我的case里GeneA的stop code和
polyA是很远的,因为中间隔了一个IRES,
问问有经验的同学,这种情况下geneA有危险么? |
|
F***s 发帖数: 12 | 4 很多人都怀疑实验室一千老的实验数据造假。大家看看下面贴的2张图:一张是投稿前1
个月的结果而另一张是投稿时的结果。这张图是文章的核心:geneA调控geneB进而调控
肿瘤细胞的生长。这张图用到了geneA的shRNA knockdown及geneB的rescue。
是不是应该将此反映给杂志社? |
|
s******y 发帖数: 28562 | 5 没有问题。
两个转染还可以同时做。根据我们转不同荧光蛋白的经验,一般来说能转进gene A 的
细胞,超过90% 也会同时转上geneB |
|
F***s 发帖数: 12 | 6 其实我觉得HA还不是最大的问题。投稿前1个月的图中2个shRNA都knockdown了geneA,但
其中一个并没有导致geneB下降,而右边的细胞生长曲线2个shRNA却是相似的。在投稿
时的图就被改过来了。
前1 |
|