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用人的周围白细胞和对温度敏感的CHO细胞系[其甲硫氨酰tRNA合成酶(MetRS)已发生改变]进行体细胞杂交,在39℃选择杂交体。这种温度对突变型CHO亲体是有害的,因此,要维持和表达补偿有缺陷的CHO基因的人类基因则需要杂交.在体外,经温度抗性杂交体的无细胞提取液的热灭活效应证明,存在两种形式的MetRS。一种与突变型CHO的酶相似,而另一种则与正常人的酶相似。用胰蛋白-姬姆萨(Giemsa)分带,碱性姬姆萨染色以及染色体同工酶标记分析5种单独的温度抗性杂交体揭示,5种杂交体都保留(retain)有12号染色体。其中3种仅有12号染色体,另外2种或
Somatic hybridization was performed with human peripheral leukocytes and with temperature-sensitive CHO cell lines [whose methionyl tRNA synthetase (MetRS) has been altered] and the hybrids were selected at 39 ° C. This temperature is detrimental to the mutated CHO parent and therefore requires hybridization in order to maintain and express the human gene that compensates for the defective CHO gene.In vitro the heat inactivation effect of the cell-free extract of the temperature-resistant hybrid Prove that there are two forms of MetRS. One is similar to the mutated CHO enzyme, while the other is similar to normal human enzymes. Analysis of five separate temperature-resistant hybrids with the Giemsa zoning, basic Giemsa staining, and chromosomal isozyme labeling revealed that all five hybrids retained chromosome 12 . Three of them are only chromosome 12, the other two or