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目的制备地高辛标记的人酪氨酸羟化酶（ｈＴＨ２）ｃＲＮＡ探针。方法用分子克隆技术重组质粒ｐＧＥＭＴＨ２。用体外转录法制备地高辛标记ｈＴＨ２ｃＲＮＡ探针，经限制性内切酶分析，显示重组质粒含有酪氨酸羟化酶基因片段，且插入位点正确。使用斑点杂交证实我们制备的探针是敏感而可靠的。结果杂交阳性部位呈紫蓝色，深浅与稀释度成正比。结论Ｄｉｇ标记的ｈＴＨ２ｃＲＮＡ探针有较好的敏感性和可靠性，可用于基因治疗帕金森病动物模型基因表达的研究。
Objective To prepare digoxigenin-labeled human tyrosine hydroxylase (hTH2) cRNA probe. Methods The recombinant plasmid pGEMTH2 was constructed by molecular cloning technique. Digoxigenin-labeled hTH2cRNA probe was prepared by in vitro transcription. The restriction endonuclease analysis showed that the recombinant plasmid contained tyrosine hydroxylase gene fragment, and the insertion site was correct. Dot blotting confirmed that the probe we prepared was sensitive and reliable. Results The positive hybridization site was purple-blue, the depth is proportional to the dilution. Conclusion Dig-labeled hTH2cRNA probe has good sensitivity and reliability and can be used for gene therapy of animal model of Parkinson’s disease.