Construction and characterization of a synthetic Baculovirus-inducible 39K promoter

被引:5
作者
Dong, Zhan-Qi [1 ]
Hu, Zhi-Gang [1 ]
Li, Hai-Qing [1 ]
Jiang, Ya-Ming [1 ]
Cao, Ming-Ya [3 ]
Chen, Peng [1 ]
Lu, Cheng [1 ,2 ]
Pan, Min-Hui [1 ,2 ]
机构
[1] Southwest Univ, State Key Lab Silkworm Genome Biol, Chongqing 400716, Peoples R China
[2] Southwest Univ, Key Lab Sericulture Funct Genom & Biotechnol, Agr Minist, Chongqing 400716, Peoples R China
[3] Henan Univ, Sch Med, Inst Immunol, Joint Natl Lab Antibody Drug Engn, Kaifeng 475004, Peoples R China
基金
中国博士后科学基金; 中国国家自然科学基金;
关键词
Baculovirus; 39; K; Inducible promoter; Synthetic; NUCLEAR POLYHEDROSIS-VIRUS; TRANSGENE EXPRESSION; GENE; SILKWORM; ESTABLISHMENT; OPTIMIZATION; SYSTEM;
D O I
10.1186/s13036-018-0121-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Background: Silkworm genetic engineering is widely used in gene function, silk engineering and disease-resistant engineering in most of Asia. Some of the earliest promoter elements are used to control the development of silkworm transgenic expression and gene therapy. However, the low expression and specificity of natural promoters limit the applications of genetic engineering. To construct a highly efficient synthetic inducible promoter in the Bombyx mori (Lepidoptera), we analyzed the regulatory elements and functional regions of the B. mori nucleopolyhedrovirus 39 K promoter. Results: Truncated mutation analysis of the 39 K promoter showed that the transcriptional regulatory region spanning positions -573 to -274 and +1 to +62 are essential for virus-inducible promoter activity. Further investigations using the electrophoretic mobility shift assay revealed that the baculovirus IE-1 protein binds to the 39 K promoter at the -310 to -355 region, and transcription activates the expression of 39 K promoter assay. Finally, we successfully constructed a synthetic inducible promoter that increased the virus-inducing activity of other promoters using the baculovirus-inducible transcriptional activation region that binds to specific core elements of 39 K (i.e., spanning the region -310 to -355). Conclusions: In summary, we constructed a novel, synthetic, and highly efficient biological tool, namely, a virus-inducible 39 K promoter, which provides endless possibilities for future research on gene function, gene therapy, and pest control in genetic engineering.
引用
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页数:12
相关论文
共 42 条
[1]   Tuning genetic control through promoter engineering [J].
Alper, H ;
Fischer, C ;
Nevoigt, E ;
Stephanopoulos, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (36) :12678-12683
[2]   Screening and optimization of an efficient Bombyx mori nucleopolyhedrovirus inducible promoter [J].
Cao, Ming-ya ;
Kuang, Xiu-xiu ;
Li, Hai-qing ;
Lei, Xue-jiao ;
Xiao, Wen-fu ;
Dong, Zhan-qi ;
Zhang, Jun ;
Hu, Nan ;
Chen, Ting-Ting ;
Lu, Cheng ;
Pan, Min-Hui .
JOURNAL OF BIOTECHNOLOGY, 2016, 231 :72-80
[3]   FUNCTIONAL MAPPING OF AN ACNPV IMMEDIATELY EARLY GENE WHICH AUGMENTS EXPRESSION OF THE IE-1 TRANS-ACTIVATED 39K-GENE [J].
CARSON, DD ;
GUARINO, LA ;
SUMMERS, MD .
VIROLOGY, 1988, 162 (02) :444-451
[4]   In vivo characterization of plant promoter element interaction using synthetic promoters [J].
Cazzonelli, Christopher Ian ;
Velten, Jeff .
TRANSGENIC RESEARCH, 2008, 17 (03) :437-457
[5]  
CHKONIYA T T, 1991, Molekulyarnaya Biologiya (Moscow), V25, P1427
[6]   EXPRESSION OF THE IE1 TRANSACTIVATOR OF AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS DURING VIRAL-INFECTION [J].
CHOI, J ;
GUARINO, LA .
VIROLOGY, 1995, 209 (01) :99-107
[7]   Promoter and Terminator Discovery and Engineering [J].
Deaner, Matthew ;
Alper, Hal S. .
SYNTHETIC BIOLOGY - METABOLIC ENGINEERING, 2018, 162 :21-44
[8]   Establishment of a highly efficient virus-inducible CRISPR/Cas9 system in insect cells [J].
Dong, Zhan-Qi ;
Chen, Ting-Ting ;
Zhang, Jun ;
Hu, Nan ;
Cao, Ming-Ya ;
Dong, Fei-Fan ;
Jiang, Ya-Ming ;
Chen, Peng ;
Lu, Cheng ;
Pan, Min-Hui .
ANTIVIRAL RESEARCH, 2016, 130 :50-57
[9]   Bombyx mori nucleopolyhedrovirus ORF79 is a per os infectivity factor associated with the PIF complex [J].
Dong, Zhan-Qi ;
Zhang, Jun ;
Chen, Xue-Mei ;
He, Qian ;
Cao, Ming-Ya ;
Wang, La ;
Li, Hai-Qing ;
Xiao, Wen-Fu ;
Pan, Cai-Xia ;
Lu, Cheng ;
Pan, Min-Hui .
VIRUS RESEARCH, 2014, 184 :62-70
[10]  
FRIESEN PD, 1986, CURR TOP MICROBIOL, V131, P31