人CD23基因的扩增及其在HEK 293T细胞的高效表达

目的建立一个有效表达CD23的系统。方法采用RT-PCR方法,从外周血淋巴细胞中扩增人的CD23cDNA基因,并将其克隆到真核表达载体pcDNA3·1B上,构建成pcDNA3·1/CD23质粒表达载体;将pcDNA3·1/CD23质粒转染HEK293T细胞,通过流式细胞仪和Westernblot检测CD23在细胞中的表达情况。结果扩增的CD23cDNA序列与文献报道的一致;通过构建表达载体和转染细胞实现了CD23cDNA在293T细胞膜上的表达。转染效率达80%以上,并获得了较高水平的表达。结论扩增CD23cDNA基因,经过转染HEK293T细胞,实现了CD23在HEK293T细胞的高效表达。

Amplification of human CD23 gene and its high-level expression in HEK293T cells

Objective To establish an efficient CD23 expression system. Methods Human CD23 cDNA was amplified by RT-PCR from peripheral lymphocytes and then inserted into pcDNA3.1A expression vector to construct the pcDNA3.1/CD23 recombinant plasmid. The pcDNA3.1/CD23 plasmids were transfected into HEK293T cells and the expressed CD23 protein was detected by flow cytometry and Western blot. Results The results showed that the sequence of CD23 cDNA amplified was consistent with that previously reported. The highly efficient expression of CD23 in HEK293T cells was achieved after transfection. The high-efficiency transfection (above 80%) and high-level expression were obtained. Conclusion Human CD23 cDNA was amplified and the high-level expression of CD23 in 293T cells was achieved.