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将大鼠脑原生型一氧化氮合酶（cNOS）全长cDNA定向插入pRC／CMVpolylinker区，获得真核细胞表达载体pCMVcNOS。以pCMVcNOS为模板，cNOS内部基因序列设计的引物进行PCR扩增，证明cNOS基因的存在。经酶切检测进一步证实插入方向完全正确。用磷酸钙共沉淀法和LipofectinTM法将pCMVcNOS转染NG108-15细胞，用含600mg／LG418培养基筛选和增殖抗性单克隆细胞，通过3H-精氨酸转化3H-胍氨酸法测定各细胞克隆可溶相和颗粒相cNOS活性，筛选高效稳定表达的细胞株。从42株抗G418单克隆细胞林中筛选到3株稳定高效表达细胞，在表达的cNOS活性中，可溶相增加更为明显。本实验结果证实，得到高效表达原生型一氧化氮合酶的神经细胞株。
The cNOS full-length cDNA was inserted into the pRC/CMV polylinker region and the eukaryotic cell expression vector pCMVcNOS was obtained. Using pCMVcNOS as a template and the primers designed for the internal gene sequence of cNOS for PCR amplification, the existence of cNOS gene was confirmed. Enzymatic detection further confirmed that the insertion direction was completely correct. pCMVcNOS was transfected into NG108-15 cells by calcium phosphate co-precipitation method and LipofectinTM method. Resistant monoclonal cells were screened and proliferated with 600 mg/LG418 medium, and 3H-proline was used to transform 3H-proline. Clone soluble and granular cNOS activity was cloned and efficient and stable cell lines were screened. Three stable and high expressing cells were screened from 42 strains of anti-G418 monoclonal cell lines. The soluble phase was more evident in the expressed cNOS activity. The results of this experiment confirmed that a neuronal cell line that expresses native nitric oxide synthase highly efficiently was obtained.