I uesd the pLKO AS2 as my lentiviral vector. Visit My Website
I inserted 750-bp genes of interest into the vecors. The processes of lentiviral production and infection seem successful because my infected cells (LNCaP, a prostate cancer cell line) survive well after G418 selection(about 3 days). However, no exogenous protein was expressed when I checked the expression level with Western blot!
It seems that my genes in constructs have frame shifts. But it is little possible because all of 4 constructs have the same failure results! Could it happen even when I used Pfu polymerases to amplifiy my genes?! I inspected my primers and they are right! Anyway, I will sequence all the constructs again!
This awesome troulble almost killed me! Plz would anyone who has done this technique give me a little suggestions? Thanks!
Edited by icome, 20 March 2010 - 01:46 AM.