Hemagglutinin-neuraminidase (HN) protein was expressed in COS-7 cells, indicating that the expression of HN protein driven by SRα promoter is higher than that driven by chicken β-actin promoter. Moreover, with 5'...Hemagglutinin-neuraminidase (HN) protein was expressed in COS-7 cells, indicating that the expression of HN protein driven by SRα promoter is higher than that driven by chicken β-actin promoter. Moreover, with 5' noncod-ing region (NCR) of HN gene, the expression was enhanced. Northern blotting demonstrated that this phenomenon was caused by the difference of HN mRNA transcription. To know the regulatory function of 5' NCR, HN gene 5' NCR was replaced by 5' NCR of keratin gene or cytochrome P-450 gene and the 3' NCR was deleted by site-directed mutagenesis. By using CAT gene as a reporter, SI nuclease assay was done to quantitate the HN mRNA transcript in the COS-7 cells co-transfected with the reporter and mutated plasmids, indicating that 5' NCR is non-specific to the enhancement of HN protein expression, and the 3' NCR also has a special regulatory function.展开更多
文摘Hemagglutinin-neuraminidase (HN) protein was expressed in COS-7 cells, indicating that the expression of HN protein driven by SRα promoter is higher than that driven by chicken β-actin promoter. Moreover, with 5' noncod-ing region (NCR) of HN gene, the expression was enhanced. Northern blotting demonstrated that this phenomenon was caused by the difference of HN mRNA transcription. To know the regulatory function of 5' NCR, HN gene 5' NCR was replaced by 5' NCR of keratin gene or cytochrome P-450 gene and the 3' NCR was deleted by site-directed mutagenesis. By using CAT gene as a reporter, SI nuclease assay was done to quantitate the HN mRNA transcript in the COS-7 cells co-transfected with the reporter and mutated plasmids, indicating that 5' NCR is non-specific to the enhancement of HN protein expression, and the 3' NCR also has a special regulatory function.
