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将含有ＮｅｏＲ基因的ｐＳＶ２－ｎｅｏ质粒经扩增、提取及纯化，以磷酸钙共沉淀法转染入ＭＤＡ人乳腺癌细胞系，经Ｇ４１８长期高压筛选出７株抗性ＭＤＡ细胞克隆，采用ＮｅｏＲ基因特异性引物进行聚合酶链反应，证实了ＮｅｏＲ基因在各株Ｇ４１８抗性ＭＤＡ细胞基因组中的整合。ＭＤＡ－ＮｅｏＲ细胞在传代６个月之后仍可耐受６００ｍｇ／Ｌ高浓度的Ｇ４１８，说明ＮｅｏＲ基因导入后的整合及表达是稳定的。表明已初步建立了携有特异标志基因ＮｅｏＲ的人乳腺癌细胞模型。
The pSV2-neo plasmid containing NeoR gene was amplified, extracted and purified, and then transfected into MDA human breast cancer cell line by calcium phosphate co-precipitation method. Seven resistant MDA cell clones were screened by G418 long-term high pressure. NeoR gene Specific primers were used for polymerase chain reaction to confirm the integration of the NeoR gene in the genome of each strain of G418-resistant MDA. MDA-NeoR cells can tolerate 600mg / L of G418 after 6 months of passage, indicating that the integration and expression of NeoR gene after its induction are stable. This indicates that a human breast cancer cell model bearing the specific marker NeoR has been initially established.