用中国仓鼠卵巢细胞系进行染色体介导的基因转移。

Chromosome-mediated gene transfer with the Chinese hamster ovary cell line.

作者信息

Lewis W H

出版信息

Exp Cell Res. 1983 Feb;143(2):309-18. doi: 10.1016/0014-4827(83)90056-3.

Abstract

Using an improved method of chromosome-mediated gene transfer, we have investigated transfer of the codominantly expressed methotrexate-resistant dihydrofolate reductase (MtxRIIIdhfr) gene into Chinese hamster ovary (CHO) cell recipients. The frequency of dhfr gene transfer with CHO cells varied considerably from clone to clone, ranging from 4 X 10(-7) to 5 X 10(-5). Using appropriate cell recipients we were able to test for linkage of several genetic markers available in the CHO cell line. For example, the mutation resulting in the auxotrophic glyB-CHO cell line has been reported by others to be linked to the dhfr gene. However, we could not demonstrate cotransfer of these two markers when glyB- recipient cells were treated with MtxRIII chromosomes and transformant clones were selected for either methotrexate-resistance (MtxR) or glycine prototrophy. We conclude that these two genes are not closely linked in the hamster genome. However, the genes for thymidine kinase (tk) and galactokinase (gk), which are known to be linked in mammalian genomes, were found to cotransfer into CHO recipients with a frequency of about 50%.

摘要

利用一种改进的染色体介导的基因转移方法，我们研究了共显性表达的甲氨蝶呤抗性二氢叶酸还原酶（MtxRIIIdhfr）基因向中国仓鼠卵巢（CHO）细胞受体的转移。用CHO细胞进行dhfr基因转移的频率在不同克隆之间差异很大，范围从4×10⁻⁷到5×10⁻⁵。使用合适的细胞受体，我们能够检测CHO细胞系中几种可用遗传标记的连锁情况。例如，导致营养缺陷型glyB-CHO细胞系的突变，其他人已报道其与dhfr基因连锁。然而，当用MtxRIII染色体处理glyB⁻受体细胞并选择对甲氨蝶呤抗性（MtxR）或甘氨酸原养型的转化克隆时，我们无法证明这两个标记的共转移。我们得出结论，这两个基因在仓鼠基因组中并非紧密连锁。然而，已知在哺乳动物基因组中连锁的胸苷激酶（tk）和半乳糖激酶（gk）基因，被发现以约50%的频率共转移到CHO受体中。