To Compare NeuN Sequence In Different Strains

NeuN is a neuron-specific nuclear protein. It expresses in the majority of nervecells in the vertebrate rat nervous system. NeuN is a soluble protein and shows twobands of46kD and48kD using Western blot. It can bind DNA and RNA in vitro.Both in vivo and in vitro, lots of researches demonstrated that NeuN is aneuronal-specific marker for neurons which are differentiation and maturation. Kim etal. in2009considered NeuN is similar to Fox-3,which belongs to Fox-1gene familyowning transcription, alternative splicing function. It is a milestone for further studingNeuN.Objective: This study was designed to investigate the cloning of the neuron-specificnuclear protein NeuN from the brain tissues of mouse, rat and human. And for thepurpose to find the similarities and differences among the three strains, we sequencethe plasmids containing NeuN from different strains and compare them throughanalyzing the sequences. Based on this, we can further explore and study themolecular characteristics of NeuN and the specific mechanisms of its development,differentiation and functional differentiation in nervous system. It’s also provide thebasis for future experiments about how to select different antibodies associatedstrains.Material and Method: Total RNA were extracted from fresh brain tissues formouse and rat, and human brain tissue from paraffin-embedded sections. The reversetranscription kit was used to obtain cDNA, and the NeuN gene was amplified withRT-PCR technology. Then the PCR product was inserted into the pcDNA vector andsequenced them as well as compared with each other.Results:1. After the amplying the target genes with RT-PCR, the results shows that shows that there was a specific band near1000bp through gel electrophoresis. Thisband was thought to be NeuN.2. The results shows that all samples have obviously bands after the PCR product wasinserted into plasmids through gel electrophoresis. It meets the requirements of thetest.3. The PCR products are similar to the sequence of Fox-3gene through sequence andcomparison.4. Comparing the sequence of three different strains of NeuN using Gene Runnersoftware, the results show that about44%are conserved sequence, while56%arehighly variable sequence.Conclusion: The experiment was to get the NeuN sequence, including mouse, rat andhuman. The results showed that about44%were highly conserved sequences amongthree different strains, while56%are highly variable sequences through comparingthe sequence of three different strains of NeuN using Gene Runner software. Itsuggests that the differences of functions among the different strains may be highlyrelated with the variable sequences.