Lovering K E, Deacon N J, Mickelson C A, Vaughan H A, Curnow K M, McKenzie I F, Sandrin M S
Immunogenetics. 1987;25(6):391-6. doi: 10.1007/BF00396105.
Using DNA from L cells which expressed high levels of the CD7 (Leu-9 or HuLy-m2) antigen obtained after two cycles of transfection, a genomic library was constructed in the lambda phage Charon 4A. Recombinant clones containing the gene coding for this antigen were identified by first screening the library with both the HSV-tk gene and a probe detecting the human repetitive (Alu) sequences. DNA from 10 tk+ and 12 Alu+ recombinant clones was used to transfect L cells which were analyzed for the cell-surface expression of CD7 either early (48-72 h posttransfection) or later when hypoxanthine aminopterin thymidine-resistant colonies were obtained. Transfection with either Alu+ or tk+ recombinant phages led to transient early expression of CD7, and stable CD7+ transfectants were also established. Thus the CD7 gene has been isolated in a number of clones in association with either the Alu repetitive sequence or with the HSV-tk gene; the insert size in one of the genomic clones was 13.5 kb.
利用经过两轮转染后获得的、表达高水平CD7(Leu-9或HuLy-m2)抗原的L细胞的DNA,构建了λ噬菌体Charon 4A基因组文库。通过先用单纯疱疹病毒胸苷激酶(HSV-tk)基因和检测人重复(Alu)序列的探针筛选文库,鉴定出含有该抗原编码基因的重组克隆。来自10个tk +和12个Alu +重组克隆的DNA用于转染L细胞,在转染早期(转染后48 - 72小时)或之后获得次黄嘌呤氨基蝶呤胸苷抗性菌落时,对L细胞进行CD7细胞表面表达分析。用Alu +或tk +重组噬菌体转染导致CD7的瞬时早期表达,并且还建立了稳定的CD7 +转染细胞系。因此,CD7基因已在多个与Alu重复序列或HSV-tk基因相关的克隆中分离出来;其中一个基因组克隆中的插入片段大小为13.5 kb。