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Modification of pGH cDNA using the first intron and adenovirus-mediated expression in CHO cells
Xiujin Li1, Fei Zhong, Shunzhang Qi
1Department of Biotechnology, Yanshan University, Qinhuangdao 066004, China.
Objective:
This study was conducted to investigate the function of the first intron of porcine growth hormone (pGH) gene in the gene expression.
Methods:
PCR method was used to amplify the first intron from pig genomic DNA. The intron was then inserted into pGH cDNA to construct pGH cDNA-intron (pGH cDNA-in). The recombinant adenoviruses containing pGH cDNA and pGH cDNA-in genes under control of CMV promoter were generated by homologous recombination method in HEK 293 cells respectively. The effect of the first intron on gene expression was evaluated by comparing the expression levels of pGH cDNA-in and pGH cDNA mediated by adenovirus vectors in CHO cells.
Results:
The expression level of pGH cDNA containing the first intron increased by 117%, which was significantly higher than that of pGH cDNA without the intron (P < 0.001).
Conclusion:
The first intron of pGH gene has the function to improve pGH gene expression.