The supemate of cell culture was collected 48 hours after transfection . this supernate was also called conditional media
2.
The supernate of virus culture was used as templet in overlapping pcr , then the interesting gene was obtained
3.
Multicopy integrants were screened with g418 from pichia pastoris which contains recombinant plasmid , and induced with methanol to secrete interesting peptide . the supernate of pichia pastoris culture was analysed by sds - page and western blotting . a reactive band , which the apparent molecular weight is 36kd , can be detected with sheep anti - hcmv polyclonal antibodies
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