3 replies to this topic

[niraj_262003]

Dear all,
                    Can anyone tell me a method to over express human protein in mice in-vivo. The biggest problem with my protein is it is highly unstable at physiological conditions i.e. half time is 5 min in plasma. We tried with pcDNA and pLive but we are not able to get an good expression. my protein expressed in liver that is another information i want to share with you.

Sincerely yours
Niraj

[Fizban]

niraj_262003, on Oct 16 2009, 04:31 PM, said:

Dear all,
                    Can anyone tell me a method to over express human protein in mice in-vivo. The biggest problem with my protein is it is highly unstable at physiological conditions i.e. half time is 5 min in plasma. We tried with pcDNA and pLive but we are not able to get an good expression. my protein expressed in liver that is another information i want to share with you.

Sincerely yours
Niraj

if the protein is not stable changing the vector won't affect it so much. if you know the rason of the short half life you can work on that. if you presume that ubiquitination is the reason for instability try to block it by mutation.
it's not like having the wt protein but if u want overexpression you're already outside physiological conditions
Fiz

[scolix]

Use recombinant viruses to infect the liver. I know that AAV could do it. Use a weak promoter and may be you are lucky.

[cupidstunt]

career protein?