Signal peptide of human nerve growth factor is better than signal peptide of mouse nerve growth factor in mediating secretory expression of beta-endorphin
Journal Title: Di-er Junyi Daxue Xuebao - Year 2009, Vol 30, Issue 4
Abstract
Objective:To clarify whether the signal peptide of human nerve growth factor can mediate secretory expression of beta-endorphin and whether there is difference between the efficiency of signal peptides from human and mouse nerve growth factor. Methods: Two kinds of eukaryotic vectors containing human or mouse signal sequence-mediated secretory expression of beta-endorphin were constructed.The culture supernatant and cells were collected 48 h after NIH3T3 cells were transfected by the two kinds of vectors, and the cover slips with single-layer cells was prepared. The concentration of beta-endorphin in the culture was determined by radio-immunoassay. The total RNA was extracted from cells and mRNA from fusion genes was assayed by RT-PCR. Cells on cover slips were subjected to immunofluorescence staining. Results: RT-PCR showed that the fusion genes were expressed in NIH3T3 cells; the expression of beta-endorphin was mainly in the cytoplasm of NIH3T3 cells. The concentrations of beta-endorphin in the supernatants 48 h after transfection with pcDNA3.1-hEP and pcDNA3.1-mEP were (280.33±24.16) pg/ml and (191.04±7.96) pg/ml (P<0.05), respectively, and they were significantly different from that of the blank control group (P<0.01). Conclusion: The signal sequence of human nerve growth factor can mediate the secretory expression of protein and the efficacy of human signal peptide is higher than that of mouse signal peptide.
Authors and Affiliations
Xue-wu XU, Shu-jun PEI, Xue-rong MIAO, Wei-feng YU
Keywords
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