Title: Transient expression of human growth hormone genomic DNA in Bombyx mori N cells
Abstract: Objective: To study the effect of intron on the exogenous gene expression in the eukaryotic cells. Methods: A full length human growth hormone(hGH) genomic DNA fragment with 4 introns and the signal peptide sequence (1.5 kb) was cloned into the eukaryotic gene expressing vector pcDNA3.0, then the vector was transfected to the Bombyx mori N(BmN) cells with lipofectin. The cell total RNA was extracted and the hGH cDNA was amplified with RT PCR.The supernatant was collected at different time points. After cloning of 3 distinct fragments of hGH into expression vector,the difference among the 3 was measured. Results: The electrophoretic analysis and immunoblot of the recombinant protein showed that the transfected cell could synthesize and excrete the peptide effectively. ELISA assay found that of the 3 fragments,the expression efficacy of full length gene was higher than the others 2, and fragment lacking the first intron had the lowest expression. Conclusion: Our data indicate that in silkworm cells the human full length gene carrying the introns and signal peptide can be expressed,indicating that introns may have an important effect in the gene expression in body.
Publication Year: 2003
Publication Date: 2003-01-01
Language: en
Type: article
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