由买买提看人间百态

boards

本页内容为未名空间相应帖子的节选和存档,一周内的贴子最多显示50字,超过一周显示500字 访问原贴
Biology版 - TLR4-EGFP 不亮
相关主题
Re: How long does it take for expression向大家请教transfection的问题
need help on Macrophage RAW 264.7 cellsAbout establishing stable transfected cell line
IRES和GFP中间放点序列,应该不影响吧请教Dual Luciferase Assay
问knockin或转基因Help with one atrial fibrillation mutation
非完整质粒DNA transfectiontransfection and cell cycle synchronization
Re: 细胞长得太密会不会影响transfected protein的表达Question: If I wanna perform flow-cytometry to check function of a gene...
Arguing with Boss with co-transfection求助Hela transfection
MLE-15 transfectiontransfection neural stem cell
相关话题的讨论汇总
话题: tlr4话题: egfp话题: 不亮话题: pegfp话题: n1
进入Biology版参与讨论
1 (共1页)
p******i
发帖数: 1092
1
我把TLR4 cDNA (2.6kb)从macrophage里PCR出来,加上XhoI和BamHI,接到
pEGFP-N1里,测序结果也是正确的,就是TLR4后面接了个in frame fusion的EGFP
但问题是transfect到293T里不亮,而空载体pEGFP-N1很亮……
我现在怀疑是kozak不够强,准备在ATG前面加一个
GCCACCATGGXX,不知道会不会有帮助……
哪位做过类似的实验,请不吝赐教啊,
在下先谢过了……
i**********a
发帖数: 1402
2
stop codon 去掉了吗
m***c
发帖数: 637
3
i did a 3kb cdna-GFP fusion protein, signal is very weak, it may hide the
green color
C*******e
发帖数: 4348
4
加linker了么?
会不会后面EGFP没折叠好
做了别的实验验证的确有EGFP fusion protein表达么?
1 (共1页)
进入Biology版参与讨论
相关主题
transfection neural stem cell非完整质粒DNA transfection
CRISPR/CAS9为啥要用virus delivery system呢?Re: 细胞长得太密会不会影响transfected protein的表达
急求一篇paper!!! thanks!Arguing with Boss with co-transfection
这句话是什么意思 帮忙解释一下~MLE-15 transfection
Re: How long does it take for expression向大家请教transfection的问题
need help on Macrophage RAW 264.7 cellsAbout establishing stable transfected cell line
IRES和GFP中间放点序列,应该不影响吧请教Dual Luciferase Assay
问knockin或转基因Help with one atrial fibrillation mutation
相关话题的讨论汇总
话题: tlr4话题: egfp话题: 不亮话题: pegfp话题: n1