Cloning of the Mouse IL-23 Gene and Its Expression in Retrovirus

To construct retroviral vector for mouse IL-23 gene, total DNA was extracted from the tumor tissues containing activated lymphocytes,and the cDNA was obtained by RT-PCR,which was just the same as that was reported in GeneBank.This cDNA was inserted into retroviral vector LXSN,transfected E.coli DH5α,and expanded. Then,two packing cell lines (ecotrophic ψ2 and amphotrophic PA317) were transfected,and the positive cellular clones were screened by G418. The expression of the desired gene was determined by PCR,RT-PCR and Northern blotting analysis. By using the above mentioned methods,the mouse IL-23 cDNA had been inserted into retroviral vector successfully,and the cellular clone PA317 with high level expression of IL-23 was produced through transfections of two packing cell lines with retroviruses.