Cloning, Expression And Primary Functional Analysis Of PDIP1γ Gene

Tumor necrosis factor a (TNF- a ) is a multifunctional cytokine that plays roles in a variety of cellular processes, such as apoptosis, proliferation, B cell activation, liver regeneration and some inflammatory responses. Human polymerase delta-interacting protein 1 (PDIPl) is a sort of protein induced by TNF- a and interleukin 6(IL-6). Besides, PDIPl can interact directly with proliferating cell nuclear antigen(PCNA) and the small subunit of DNA polymerase 5 . The results of searching human genome indicate that there may be two more homolog genes respect to hPDIPl, the case is similar in the mouse genome. The homologies encode proteins of conservative amino acid sequence, each has a BTB/POZ domain in the N'-terminal and conserved PCNA-binding motif (QTKV-EFP) in the C'-terminal. Thus we rank them PDIPl gene family. To know more functions about the PDIPl family, we firstly cloned the mouse mPDIPl? gene, and expressed it in E.coli. Then the purified protein from the bacteria lysis was used to immunize the New Zealand rabbit, and we got the highly- titrate specific polyclonal antibody to mPDIPl?. We identified the expression profiles of mPDIPl? by Northern-Blot and Western-Blot respectively, which reflecting the native RNA and protein levels. Simultaneity we also cloned human hPDIPl? gene, and demonstrated that hPDIPl? interacted with cyclin-dependent inhibitor protein (p21) in vivo through co-immunoprecipitation assay. In conclution, these work will promote the functional reseach of gene family.