Nesterova T B, Isaenko A A, Matveeva N M, Shilov A G, Rubtsov N B, Vorob'eva N V, Rubtsov N V, Vandeberg J, Zakiian S M
Southwestern Center of Biomedical Research, San-Antonio 28147.
Genetika. 1994 Nov;30(11):1516-24.
A possibility of obtaining a panel of marsupial-rodent somatic cell hybrid clones has been explored, with a view to mapping the genome of the opossum (Monodelphis domestica). Fusion of opossum cells (splenocytes, bone marrow cells, and fibroblasts) with fibroblasts of Chinese hamster or vole (HGPRT- and TK- mutants, respectively) produced 146 hybrid clones. The majority of marsupial-mammalian somatic cell hybrids were characterized by pronounced fragmentation and segregation of marsupial chromosomes. To overcome this difficulty, a method for rapid screening was developed, which allowed the early selection of clones rich in chromosomal material of opossum. Based on the screening results, 25 clones of independent origin were selected. A detailed genetic analysis, which included chromosome G-banding and in situ hybridization of biotin-labeled opossum DNA on metaphase chromosomes, allowed further selection of seven hybrid clones containing one to six intact chromosomes of M. domestica. Opossum chromosomes were present in various combinations against the background of Chinese hamster or vole chromosomes. The clones will be included in the panel of opossum-rodent somatic cell hybrids, which is currently being created.
为了绘制负鼠(家短尾负鼠)的基因组图谱,人们探索了获得一组有袋类 - 啮齿类体细胞杂交克隆的可能性。将负鼠细胞(脾细胞、骨髓细胞和成纤维细胞)与中国仓鼠或田鼠的成纤维细胞(分别为次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶缺陷型和胸苷激酶缺陷型突变体)融合,产生了146个杂交克隆。大多数有袋类 - 哺乳类体细胞杂种的特征是负鼠染色体明显断裂和分离。为克服这一困难,开发了一种快速筛选方法,该方法可早期筛选出富含负鼠染色体物质的克隆。根据筛选结果,选择了25个独立来源的克隆。详细的遗传分析,包括染色体G显带和生物素标记的负鼠DNA在中期染色体上的原位杂交,进一步筛选出7个含有1至6条家短尾负鼠完整染色体的杂交克隆。在中华仓鼠或田鼠染色体的背景下,负鼠染色体以各种组合形式存在。这些克隆将被纳入目前正在创建的负鼠 - 啮齿类体细胞杂种库中。