CAJ | 학술논문

目的：克隆HLA-A*0201编码区的cDNA，并建立在COS-7细胞上的瞬时表达系统。方法：采用逆转录-PCR技术从HLA-A*0201阳性的淋巴细胞中获得HLA-A的cDNA，将其定向克隆至载体pBluescriptⅡSK(+/-)中，经测序确证后，将该基因插入真核表达载体pcDNA3中，通过阳离子脂质体介导转染至COS-7细胞，用流式细胞仪测定细胞表面HLA-A*0201分子的瞬时表达。结果：获得的HLA-A*0201基因的cDNA序列与Genebank登录的cDNA序列一致。FACS检测结果显示，COS-7细胞上HLA-A*0201的表达率为53％。结论：成功克隆HLA-A*0201基因，并实现了在COS-7细胞上的瞬时表达，为研究HLA-A2限制性的肿瘤杀伤作用奠定了基础。
목적：극륭HLA-A*0201편마구적cDNA，병건립재COS-7세포상적순시표체계통。방법：채용역전록-PCR기술종HLA-A*0201양성적림파세포중획득HLA-A적cDNA，장기정향극륭지재체pBluescriptⅡSK(+/-)중，경측서학증후，장해기인삽입진핵표체재체pcDNA3중，통과양리자지질체개도전염지COS-7세포，용류식세포의측정세포표면HLA-A*0201분자적순시표체。결과：획득적HLA-A*0201기인적cDNA서렬여Genebank등록적cDNA서렬일치。FACS검측결과현시，COS-7세포상HLA-A*0201적표체솔위53％。결론：성공극륭HLA-A*0201기인，병실현료재COS-7세포상적순시표체，위연구HLA-A2한제성적종류살상작용전정료기출。
Objective:To clone HLA-A* 0201 cDNA and express it transiently on COS-7 cells.Methods: HLA-A cDNA isolated from the HIA-A* 0201 positive lymphocytes by using RT-PCR was cloned into pBluescript Ⅱ SK vector directionally. After the sequence was confurmed,the cDNA was inserted into mammalian expression vector pcDNA3.The recombinant vector was transfected into COS-7 cells by cation liposome.The transient expression on the cells was measured by flow cyteometer. Results:The cDNA was i dentical with HLA-A* 0201 cDNA published on Genebank. Determined by flow cytemeter,the expressing rate was recorded for 57%. Conclusion: We have cloned HLA-A* 0201 successfully and expressed it transiently on COS-7 cell,which would be potentially useful in research on killing tumor restricted by HLA-A2.