表达人白细胞介素-2的重组腺病毒的制备  

Construction of Recombinant Adenovirus for Expressing Human Interleukin-2

作　　者：徐人尔[1] 应蓓蓓[1] 王义斌[2] 赵寿元[1] 李昌本[1] 

机构地区：[1]复旦大学遗传学研究所遗传工程国家重点实验室 [2]ScridppsResearchInstitute,LaJolla,CA92037,USA

出　　处：《高技术通讯》1997年第1期15-18,共4页Chinese High Technology Letters

摘　　要：携带人白细胞介素-2（interleukin－2，IL－2）cDNA序列的E1区缺陷的腺病毒载体pAdl2／RSV－IL2－bpA与pJM17质粒，通过磷酸钙沉淀法共转染293细胞，经同源重组，获得了一个腺病毒噬斑。抽提腺病毒DNA，运用PCR扩增及XbaI酶切检测，证实这一噬斑为带有人IL－2cNDA序列的重组腺病毒。受其感染的293细胞和人黑素瘤A375细胞上清中，都能够测出IL－2的活性，表明制备的重组腺病毒是能表达人IL－2的，可进一步用于恶性肿瘤的基因治疗。E1-deleted adenoviral vector, pAd12 RSV-IL- bpA, harboring human IL- 2 cDNA ,and plasmid pJM17 are co- transfected into 293cells by calium- phosphate precipitation-mediated transfection. One recombinant adenovirus (rAd) is obtained by homologous recombination after 11 days incubstion. PCR detection assay using viral DNA as template and subsequent Xbal digestion analysis indicates that this rAd containshuman IL-2 cDNA. The biological activity of IL-2 is measured from the supernatant of the cultured medium from the 293 cells and A37 cells following rAd infection.

关 键 词：人白细胞介素-2 腺病毒 重组体 

分 类 号：Q78[生物学—分子生物学]