Difference between revisions of "Part:BBa K3064008"

Latest revision as of 11:49, 20 October 2019

PGK promoter

PGK promoter is a universal promoter which has a strong activation function and it has high content of G and C. PGK promoter is more effective than CMV in some tissue cells such as striatal cells and it is universally expressed in most organizations. PGK promoter is often used as a strong promoter to initiate the transcription of exogenous genes. People insert PGK promoter into vector to drive the expression of downstream target genes.

Experiment

We inserted PGK promoter into plasmid pmirGLO and the promoter is upstream of the gene of fire luciferase.

We laid cells, HepG2, in a 24-hole plate. When the confluence reached 70-90, pmirGLO plasmid was transfected. And then, culture media were divided into six groups of sugar concentration: 0mM, 5mM, 10mM, 15mM, 20mM, 30mM and were used to stimulate cells. After that, the cells were lysed and tested with a double luciferase kit. We found that expression of luciferase had significant changes with different sugar concentration.

The extracellular sugar concentration in human physiological environment is about 22mM. And the experimental data showed that firefly luciferase gene which is downstream of PGK promoter expressed better and better when the sugar concentration is gradually approaching the physiological sugar concentration 22mM.

References

[1] Xinfeng yang,AnPing Guo,Hua Kong,et al.Cloning and functional analysis of industrial saccharomyces cerevisiae PGK promoter[J].Chinese Agricultural Science Bulletion,2012,28(06):178-182(Chinese)

[2]ZhiJun Zhou,YongKang Zou,JianGuo Li,et al.Construction and identification of lentivirus expression plasmid containing PGK promoter[J].Journal of Anhui Normal University(Natural Science),2009,32(02):163-167(Chinese)

Sequence and Features