PMID- 29027455
OWN - NLM
STAT- MEDLINE
DCOM- 20180409
LR  - 20181202
IS  - 1000-3061 (Print)
IS  - 1000-3061 (Linking)
VI  - 32
IP  - 10
DP  - 2016 Oct 25
TI  - [Construction and verification of NF-kappaB luciferase reporter gene system].
PG  - 1465-1473
LID - 10.13345/j.cjb.160185 [doi]
AB  - To quantify the transcriptional activity of NF-kappaB and to screen drugs related to 
      the regulation of NF-kappaB activation, we constructed a recombinant plasmid through 
      deleting the original CMV promoter of retrovirus vector pQCXIP and inserting the 
      NF-kappaB enhancer and NanoLuc luciferase sequence into the vector. Then, using the 
      recombinant plasmid we constructed a cell line in which the expression of NanoLuc 
      luciferase (NLuc) was regulated by NF-kappaB. The inserted sequences were verified by 
      restriction endonuclease digestion and sequencing. Tumor necrosis factor-alpha 
      (TNF-alpha), an NF-kappaB activator, acted on the constructed NLuc cell line and leaded 
      to the specific luciferase reaction. The luciferase reaction showed a fine time 
      and dose dependence to the TNF-alpha stimulation, indicating the successful 
      construction of the NF-kappaB regulated NLuc-expressing cell line. Besides, the NF-kappaB 
      inhibitor, triptolide, reduced the expression of NLuc in a dose-dependent way. 
      The constructed reporter system in this study could be applied in the 
      quantification of the NF-kappaB transcriptional activity and in the NF-kappaB 
      regulation-related drug screening.
FAU - Guo, Zhilan
AU  - Guo Z
AD  - College of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 
      100102, China.
AD  - Beijing Key Laboratory of Research of Chinese Medicine on Prevention and 
      Treatment for Major Diseases, Experimental Research Center, China Academy of 
      Chinese Medical Sciences, Beijing 100700, China.
FAU - Che, Luyang
AU  - Che L
AD  - Department of Orthopedics, Chinese People's Liberation Army General Hospital, 
      Beijing 100853, China.
FAU - Li, Jingzhe
AU  - Li J
AD  - Beijing Key Laboratory of Research of Chinese Medicine on Prevention and 
      Treatment for Major Diseases, Experimental Research Center, China Academy of 
      Chinese Medical Sciences, Beijing 100700, China.
FAU - Sun, Zhenxiao
AU  - Sun Z
AD  - College of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 
      100102, China.
FAU - Liu, Changzhen
AU  - Liu C
AD  - Beijing Key Laboratory of Research of Chinese Medicine on Prevention and 
      Treatment for Major Diseases, Experimental Research Center, China Academy of 
      Chinese Medical Sciences, Beijing 100700, China.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Sheng Wu Gong Cheng Xue Bao
JT  - Sheng wu gong cheng xue bao = Chinese journal of biotechnology
JID - 9426463
RN  - 0 (NF-kappa B)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Enhancer Elements, Genetic
MH  - Gene Expression Regulation
MH  - *Genes, Reporter
MH  - Humans
MH  - Luciferases
MH  - NF-kappa B/*genetics
MH  - Promoter Regions, Genetic
MH  - Signal Transduction
MH  - Tumor Necrosis Factor-alpha/*metabolism
OTO - NOTNLM
OT  - NF-kappaB
OT  - NanoLuc luciferase
OT  - reporter gene system
EDAT- 2017/10/14 06:00
MHDA- 2018/04/10 06:00
CRDT- 2017/10/14 06:00
PHST- 2017/10/14 06:00 [entrez]
PHST- 2017/10/14 06:00 [pubmed]
PHST- 2018/04/10 06:00 [medline]
AID - 10.13345/j.cjb.160185 [doi]
PST - ppublish
SO  - Sheng Wu Gong Cheng Xue Bao. 2016 Oct 25;32(10):1465-1473. doi: 
      10.13345/j.cjb.160185.