Construction method of human astrocytes stably expressing OXTR (oxytocin receptors)
A technology of astrocytes and oxytocin receptors, applied in botany equipment and methods, biochemical equipment and methods, using vectors to introduce foreign genetic material, etc., can solve biological events and the mechanism behind them is not clear Clarification, unclear role of oxytocin, etc.
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[0031] A method for constructing human astrocytes stably expressing oxytocin receptors, comprising the following steps:
[0032] 1) Constructing a recombinant lentiviral vector, the viral vector inserts the nucleotide sequence shown in SEQ ID NO.1 in the backbone of pCDH-CMV-MCS-EF1α-copGFP;
[0033] 2) packaging the recombinant lentiviral vector;
[0034] 3) Human astrocyte culture and screening: the packaged recombinant lentivirus was used to transduce human astrocytes to obtain human astrocytes stably expressing oxytocin receptor.
[0035] Further, the Flag-OXTR (SEQ ID NO.2) and mCherry (SEQ ID NO.3) fragments were linked together by overlapping PCR.
[0036] Further, a linker fragment (SEQ ID NO.4) was added between Flag-OXTR and mCherry to obtain a Flag-OXTR-mCherry fragment (SEQ ID NO.1),
[0037] Further, recognition sites for XbaI and EcoRI restriction endonucleases were respectively introduced at the 5' end and 3' end of the Flag-OXTR-mCherry fragment.
[0038] Fu...
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