Shupp Byrne D E, Church R L
Somatic Cell Genet. 1983 May;9(3):313-31. doi: 10.1007/BF01539141.
Somatic cell hybrids between mouse and Chinese hamster fibroblasts have been used to identify the chromosome responsible for the synthesis of both mouse type I procollagen subunit chains (MCOLA1 and MCOLA2). Thirty-one separate hybrid clones and subclones from ten separate hybridization events were isolated in hypoxanthine-aminopterin-thymidine (HAT) selection medium and were used for detailed gene-mapping studies. ELISA and "Western blotting" immunochemical analysis were used to detect the production of mouse type I procollagen in each hybrid clone. Mouse and Chinese hamster chromosomes were identified in each hybrid clone by trypsin-Giemsa banding of metaphase chromosome spreads and by isozyme analysis. We have found that mouse type I procollagen production segregates concordantly with mouse superoxide dismutase-1, previously mapped to mouse chromosome 16, and with the presence of mouse chromosome 16 karyotypically. Western blotting immunochemical analysis of the separated mouse procollagen chains produced by each hybrid line demonstrated that apparently the genes for both subunit chains are located on the same chromosome. These studies, therefore, assign the structural genes for mouse type I procollagen pro alpha 1 (MCOLA1) and pro alpha 2 (MCOLA2) chains to mouse chromosome 16.
小鼠和中国仓鼠成纤维细胞之间的体细胞杂种已被用于鉴定负责合成小鼠I型前胶原亚基链(MCOLA1和MCOLA2)的染色体。在次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷(HAT)选择培养基中分离出了来自10次独立杂交事件的31个单独的杂交克隆和亚克隆,并将其用于详细的基因定位研究。采用酶联免疫吸附测定(ELISA)和“蛋白质免疫印迹法”免疫化学分析来检测每个杂交克隆中小鼠I型前胶原的产生。通过中期染色体铺片的胰蛋白酶-吉姆萨染色带型分析和同工酶分析,鉴定了每个杂交克隆中的小鼠和中国仓鼠染色体。我们发现,小鼠I型前胶原的产生与先前定位于小鼠第16号染色体的小鼠超氧化物歧化酶-1以及核型上小鼠第16号染色体的存在一致分离。对每个杂交系产生的分离的小鼠前胶原链进行的蛋白质免疫印迹法免疫化学分析表明,显然两个亚基链的基因位于同一条染色体上。因此,这些研究将小鼠I型前胶原α1(MCOLA1)和α2(MCOLA2)链的结构基因定位到小鼠第16号染色体上。