Raimondi E, Balzaretti M, Moralli D, Vagnarelli P, Tredici F, Bensi M, De Carli L
Departimento di Genetica e Microbiologia A Buzzati Traverso, Pavia, Italy.
Hum Gene Ther. 1996 Jun 10;7(9):1103-9. doi: 10.1089/hum.1996.7.9-1103.
A human supernumerary minichromosome (MC), previously identified as a derivative of chromosome 9, has been introduced into Chinese hamster ovary (CHO) cells by means of cell fusion. A hybrid clone containing the MC as the only free human chromosome was isolated. A selectable marker gene (neo) inserted into a yeast artificial chromosome (YAC) has been successfully targeted to the MC centromeric DNA via co-transfection with chromosome-9-specific alpha satellite DNA. In situ hybridization and Southern blotting experiments demonstrated that the intact neo gene was integrated into the MC centromeric DNA. Studies on the clonal distribution and on the stability of the MC either in the presence or in the absence of the selective agent have been carried out. The MC is susceptible to further manipulations and may thus represent a model for the construction of a large-capacity vector for somatic gene therapy.
一条先前被鉴定为9号染色体衍生物的人类额外小染色体（MC）已通过细胞融合的方式导入中国仓鼠卵巢（CHO）细胞。分离出了一个含有该MC作为唯一游离人类染色体的杂交克隆。通过与9号染色体特异性α卫星DNA共转染，成功地将插入酵母人工染色体（YAC）中的一个可选择标记基因（neo）靶向到MC着丝粒DNA上。原位杂交和Southern印迹实验表明完整的neo基因已整合到MC着丝粒DNA中。已经对MC在有或没有选择剂存在的情况下的克隆分布和稳定性进行了研究。该MC易于进行进一步操作，因此可能代表一种用于构建体细胞基因治疗大容量载体的模型。
