Cloning of fullª²length heparanase gene and expression of functional heparanase in mammalian cells

    LI Yanª²Jie, LI Jiª²Xue, PENG Qingª²Zhong, TANG Guoª²Ying, LIN Yanª²Li, GUO Ruiª²Feng, ZHU Houª²Chu

    1Medical Experimental Center, Zhengzhou University, Zhengzhou 450052, China, 2Biology Engineering Institute, Academy of Military Medical Sciences, Beijing 100071, China

    ¡¾Abstract¡¿ AIM: To explore the expression character of heparanase by isolating the full encoded gene and expressing the functional enzyme in mammalian cells. METHODS: We isolated the full encoded heparanase gene from human placenta cDNA library, cloned the gene into eukarytic expression vector pCDNA4/Mycª²His, transfected it into COSª²7 cells and CHO cells and then screened the CHO cells which permanently expressed heparanase in the selected medium. RESULTS: The activity of heparanase in the mammalian cell lysate detected was much higher than that in the culture medium. The molecular mass of the heparanase protein expressed in the transfected COSª²7 and CHO cells was about 53¡Á103 (Mr) by western blot. CONCLUSION: The fullª²length heparanase gene is isolated. The functional heparanase is expressed within COSª²7 and CHO cells and few heparanse is secreted into medium. The positive CHO cells expressing active heparanase is screened, which will help explore its function and screen its antibody and antagonists from a variety of sources. ......