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| Improving Gene Expression by Introducing Upstream Frameshift and Stop Codon |
| CHEN Bin1,2, LI Jian-bin2,3, MI Zhi-qiang2, AN Xiao-ping2, LI Cun2, LIU Da-bin2, JIANG Huan-huan2, WANG Juan2, HUANG Fen2, ZHANG Bao-zhong2, FAN Hua-hao2, HE Hou-jun1, TONG Yi-gang2 |
1. College of Animal Science & Technology,Jiangxi Agricultural University,Nanchang 330045,China;
2. State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology, Academy of Military Medical Science,Beijing 100071,China;
3. College of Food Science,South China Agricultural University,Guangzhou 510642,China |
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Abstract Objectives:To improve the expression of gene of interest in a lentiviral vector with complicated gene distribution. Methods:Lentiviruses have complicated gene distributions such as overlapping Env,Rre,Rev and Tat on their genomes. To avoid damaging the reproduction efficiency of these vectors,it is better to retain as much as possible the original gene structure when replacing the unwanted Env gene with a gene of interest. A reporter gene luc was first inserted into the lentiviral vector in place of the Env gene. The resultant plasmid did not express detectable luciferase activity. In order to improve the target gene expression, a frameshift mutation and a stop codon upstream the luc gene were introduced. Results:After the introduction of the upstream frameshift mutation and a stop codon,the luciferase expression was greatly enhanced. Conclusion:By introducing frameshift mutation and stop codon in the upstream reading frame, the expression of the downstream open reading frame could be largely increased.
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Received: 03 February 2011
Published: 25 August 2011
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Cite this article:
CHEN Bin, LI Jian-bin, MI Zhi-qiang, AN Xiao-ping, LI Cun, LIU Da-bin, JIANG Huan-huan, WANG Juan, HUANG Fen, ZHANG Bao-zhong, FAN Hua-hao, HE Hou-jun, TONG Yi-gang. Improving Gene Expression by Introducing Upstream Frameshift and Stop Codon. China Biotechnology, 2011, 31(8): 92-96.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I8/92
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