Kioussis D, Wilson F, Daniels C, Leveton C, Taverne J, Playfair J H
EMBO J. 1987 Feb;6(2):355-61. doi: 10.1002/j.1460-2075.1987.tb04762.x.
A cosmid vector carrying the Epstein-Barr virus origin of replication, the EBNA-1 gene, the hygromycin phosphotransferase (hph) gene and pBR322 sequences has been constructed. This cosmid can replicate autonomously in the nucleus of human tissue culture cells, even when it carries a 35-kb long insert. The cosmid can be rescued from the transfected cells by cloning it directly into ampicillin-sensitive Escherichia coli. A gene for human tumour necrosis factor (TNF) cloned into this cosmid vector was introduced in tissue culture cells, where it was transcribed into mature mRNA.
构建了一种黏粒载体，其携带爱泼斯坦-巴尔病毒复制起点、EBNA-1基因、潮霉素磷酸转移酶（hph）基因和pBR322序列。即使携带一个35kb长的插入片段，这种黏粒也能在人组织培养细胞核中自主复制。通过将其直接克隆到对氨苄青霉素敏感的大肠杆菌中，可从转染细胞中拯救出该黏粒。克隆到这种黏粒载体中的人肿瘤坏死因子（TNF）基因被导入组织培养细胞，在那里它被转录成成熟的mRNA。
