4 replies to this topic

[clwop]

i want to express a enzyme gene about 3500bp (fusing two subunits) from Corynebacterium glutamicum in E. coli. It is almost twice as big as the enzymes i usually deal with. so i wonder if its size will cause some trouble.

Edited by clwop, 27 December 2009 - 05:41 PM.

[Vini]

No, the size  will not be a problem.....people express proteins as big as 400 kDa from E. coli............so u dont have to worry....

[clwop]

DRN, on Dec 28 2009, 11:16 AM, said:

No, the size  will not be a problem.....people express proteins as big as 400 kDa from E. coli............so u dont have to worry....

thx

[Prep!]

clwop, on Dec 28 2009, 01:12 PM, said:

DRN, on Dec 28 2009, 11:16 AM, said:

No, the size  will not be a problem.....people express proteins as big as 400 kDa from E. coli............so u dont have to worry....

thx

http://mcl1.ncifcrf....c-Esposito.html

please refer here... it may not be impossible.. but expressing large mol wt protein in E Coli may be tricky...

[Vini]

protein expression can be tricky for varied reasons.................not just the size............dont worry, we routinely express such proteins in our lab.............they may be unstable but take precautions like maintaining 4 degree, adding protease inhibitors if required and dont make it undergo many freeze-thaw cycles..............

Edited by DRN, 28 December 2009 - 12:32 AM.