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Updated: Oct 5, 2026

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
Published on: May 3, 2018
[The function of 5'flanking region of mouse neuronal CDC2-like kinase regulatory subunit p35]
1Department of Medical Genetics, the Second Military Medical University, Shanghai 200433 P. R. China.
Objective:
To clarify the possible mechanisms which control p35(nck5a) neural-specific expression.
Methods:
Thirteen luciferase expression vectors which contain different parts of 5'flanking sequence were constructed. Transient expression assay, DNase 1 hypersensitive site(DHSS) assay and primer extension assay were made to reveal the possible mechanisms of p35(nck5a) expression control.
Results:
It was found that 5'flanking sequence of p35(nck5a) contained 5455bp and there were a few possible promoters in the fragment. The result of transient expression assay in primary cerebrum cortex cell of rat (Neuron) and established non-neuron cell such as HeLa showed the loss of neural-specific expression of all the luciferase expression vectors. The result of DNase 1 hypersensitive site assay revealed a DHSS located at the -400bp upstream of p35(nck5a) in brain but not in liver. The primer extension assay found the putative transcription initiation site near the position of DHSS.
Conclusion:
The results above suggest that there is no cis-element to control the neural-specific expression in the 5'flanking sequence, and the control on the chromosome level may be critical to the neural-specific expression of p35(nck5a).
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