The Linker in pEGFP-C1 fusion protein

[songcc at 263.NET]

Do you have the experience of GFP fusion protein that doesn't work as designed. I have make a GFP-Id1 fusion protein but I fail to get the fluorescent fusion protein when I using EcoRI R.E. insert Id1 into pEGFP-C1, and transfect to Hela and MEL cell. The sequence is right. I delete the Id1 to only 50a.a. I still can not see any fluorescent. But I make ubiquitin GFP fusion protein and I can get the fluorescent. So I think the linker between the GFP and some target protein is quite important, but I do not know how long it will work.   Would you mind share the experience of make fusion protein work, and tell me more information about the fail and success design. Is there any one make a GFP fusion protein using the Clontech pEGFP-C1 again, and insert a linker between the GFP and the target protein to keep the GFP have fluorescent? 
 Look forward for any advice of you!

Song Changcheng(songcc at 263.net)
 
Institute of Cell Biology
Beijing Normal University
China, 100875


_____________________________________________
Ê׶¼ÔÚÏß--ÏȽøÖйúÈ˵ÄÍøÉϼÒÔ° http://www.263.net
Ãâ·ÑÓÊÏä ÓʼþÔÓÖ¾ Ç©ÃûÓʼþ Óʼþ¼ÓÃÜ Óʼþ×·Éíºô
ËÑË÷ÒýÇæ ¸öÈËÕ¾µã ÔÚÏßÓÎÏ· ÍøÉÏÁÄÌì ÍøÉϹҺÅ
½ðÈÚÍõ¹ú ÔÚÏßɱ¶¾ ÌøÔéÊг¡ Èí¼þÏÂÔØ ÐÝÏÐÓéÀÖ