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用限制性内切酶EcoRI从pKS(-)HTH_1切下全长为1.9 kb的人酪氨酸羟化酶基因,在T_4DNA连接酶的作用下连接在真核表达载体pCDNA_3的EcoR Ⅰ位点,构建成重组质粒pcD-NA_3HTH_1,该质粒转染COS-7细胞,免疫荧光组织化学染色证实酪氨酸羟化酶在其中的表达。
The full-length human tyrosine hydroxylase gene was cut from pKS (-) HTH-1 by restriction endonuclease EcoRI and ligated into the EcoR Ⅰ site of eukaryotic expression vector pCDNA_3 under the action of T_4 DNA ligase. The recombinant plasmid pcD-NA_3HTH_1 was constructed and transfected into COS-7 cells. The expression of tyrosine hydroxylase was confirmed by immunofluorescence staining.