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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (3): 18-24    DOI: 10.13523/j.cb.20150303
    
Screening of ER3 Sequences Enhanced Protein Expression Activity and Its Functional Regions Identification
CUI Cheng-cheng, BI Yan-hong, WANG Ying-ming, LI Pan, YANG Si-da, HUANG Fen, ZENG Wei-kun, JING Shen-rong
Medical Faculty, Kunming University of Science and Technology, Kunming 650500, China
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Abstract  

Objective: Using a constructed enhancer sequence detecting vector, screening sequences that could enhanced heterologous protein expression in Escherichia coli, and preliminary identified its functional domain by deletion mutants. Methods: Chloramphenicol acetyltransferase (CAT) gene and truncated human papillomavirus (HPV) major capsid protein gene L1 (named L11) were fusion expressed as a report gene; to screen enhancer-like sequence from the collected samples, and detected the enhanced activity by protein expression. The functional regions were identified by construction of deletion mutants. Results: One enhancer-like sequence was successfully screened, which enhanced 11-folds of chloramphenicol resistance; fusion protein expression level increased 2.26-folds; the functional areas are located in the 1~265bp. Conclusion: An enhancer-like sequence were successfully screened, which can improve the expression of heterologous gene in Escherichia coli.



Key wordsEnhancer-like sequence      Enhancer gene expression      Deletion mutants      Functional areas     
Received: 11 December 2014      Published: 25 March 2015
ZTFLH:  Q939.93  
Cite this article:

CUI Cheng-cheng, BI Yan-hong, WANG Ying-ming, LI Pan, YANG Si-da, HUANG Fen, ZENG Wei-kun, JING Shen-rong. Screening of ER3 Sequences Enhanced Protein Expression Activity and Its Functional Regions Identification. China Biotechnology, 2015, 35(3): 18-24.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150303     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I3/18


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