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［摘 要］目的 开展基因重组AT-Ⅲ的研究，试将人AT-Ⅲ cDNA在CHO细胞中进行表达。方法 将携带AT-Ⅲ基因的表达载体转染CHO细胞，用 G418筛选抗性克隆，SDS－PACE与Western Blot检测其表达产物；并采用生化法测定AT-Ⅲ的表达量，采用凝血酶空斑法测定表达产物的活性。结果 Western Blot分析发现转移至膜上的表达产物能与抗AT-Ⅲ单克隆抗体结合，分子量与人血浆中AT-Ⅲ一致，证明在CHO细胞中成功地表达了AT-Ⅲ，并具有天然AT-Ⅲ的生物活性。结论 获得了表达AT-Ⅲ CHO细胞株。
[ABSTRACT] Objective To study the gene recombination of AT-Ⅲ and try to express human AT-Ⅲ cDNA in CHO cells. METHODS: CHO cells were transfected with the expression vector carrying AT-Ⅲ gene. The resistant clones were screened by G418. The expression products of AT-Ⅲ were detected by SDS-PAGE and Western Blot. The expression of AT-Ⅲ was determined by biochemical method. Method to determine the activity of the expressed product. Results Western Blot analysis showed that the product transferred to the membrane could bind to anti-AT-Ⅲ monoclonal antibody with the same molecular weight as AT-Ⅲ in human plasma, which proved that AT-Ⅲ was successfully expressed in CHO cells and had natural AT -Ⅲ biological activity. Conclusion The AT-Ⅲ CHO cell line was obtained.