Expression Of Truncated Zmcdc5 In E.coli

cdc5 gene is necessary for the G2/M transition. Cells with mutations in cdc5 can not finish the G2/M transition, then die; Overexpression of CDC5 can shorten G2, the cell cycle as well. It also makes cells lessening. It was proved that cdc5 gene works in the processes of transcription,cell cycle and splicing of pre-mRNA. And CDCS can transact with other factors, such as NIPP, PRL, IL-2, FGF-2 and DLK.In the past, We cloned the cDNA of Zmcdc5 based on the studies on difference of gene expression in shoot and radicle of Maize seedling in our lab and then got the whole genome sequence by maize genomic library screening and inverse PCR.This time, using cDNA of Zmcdc5 as template,we amplify a sequence by means of PCR technology.and then ,using restrict endoenzyme and ligase,we conjunct the 0.8kb length DNA sequence in a expression vector ,pET-30a.After induction,expression and purification,we obtained a 35.4kDa truncated fusing ZmCDC5 protein which contains 267Aa (647 to 914Aa in ZmCDC5).With the purified protein ,we got its antibody and testified the antibody by means of Western blotting and dot blotting. We also detected ZmCDC5 in different tissues of maize.