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Effect of Signal Peptide on the Expression and Secretion of Hepatocyte Growth Factor in CHO
Xu Dongsheng, Chen Yun, Jin Jian*
Laboratory of Drug Design and Molecular Pharmacology, School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China
Abstract: The aim of this study is to investigate the improvement of hepatocyte growth factor (HGF) expression by screening secretory signal peptides in Chinese hamster ovary cell (CHO). In the experiments， the signal peptide of HGF itself were replaced by 5 different secreted signal peptides respectively by PCR method， then five plasmids containing different signal peptides were constructed and transformed into mammalian cell CHO， and HGF expressing CHO cells were identified. During the expression process， the mRNA levels of HGF were detected by qPCR and the cumulative HGF proteins in medium were evaluated by Western blot. Besides， Madin-Daby canine kidney (MDCK) cells scattering experiment was used to verify the biological activity of HGF. The results showed that during the expression process， secretory signal peptides had little effect on HGF transcriptional level and biological activity of HGF， but had changed the accumulation of HGF significantly in culture medium. In summary，the suitable secretory signal peptide promotes the secretion of foreign protein in CHO.
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