Shh/lmx1a signaling regulates the approach of NTN-modified RaSCs to differentiate dopaminergic neurons
A dopaminergic and signaling pathway technology, applied in the field of biomedicine, can solve the problems of increasing incidence of PD, limiting long-term treatment of Parkinson's disease, and failing to fundamentally restore normal function of the lesion site.
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[0049] 1. Cloning of Lmx1a gene and chimeric NTN gene and construction of recombinant plasmid
[0050] Using Lmx1a whole transcript cDNA (NM_138396) as a template, the primers are:
[0051] P1 up: 5'-GCGTCGACGTACCATGCTGGACGGCCTAAAG-3',
[0052] P1 down: 5'-CGGGATCCCGCTTATCAAGATGTGAAGTAAG-3',
[0053] The DNA fragments with SalI and EcoRV restriction sites were amplified by PCR, detected and analyzed by 1% agarose gel electrophoresis, and the amplified bands were consistent with the theoretical molecular weight. The results are shown in figure 1 a. The amplified fragment was inserted into pShuttle-CMV, and the recombinant clone was identified by double digestion with SalI and EcoRV. The results are shown in figure 1 b. The recombinant clones were sequenced, and the sequencing results were consistent with the theoretical results.
[0054] The above Lmx1a whole transcript cDNA (NM_138396) was produced by GeneCopoeiaTM.
[0055] Connect the NTN mature peptide gene to the art...
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