DNA-mediated transformation of N-acetylglucosaminyltransferase I activity into an enzyme deficient cell line.
DNA-mediated transformation of N-acetylglucosaminyltransferase I activity into an enzyme deficient cell line.
复制标题
DNA 介导的 N-乙酰氨基葡萄糖转移酶 I 活性转化为酶缺陷细胞系。
DOI：
10.1016/0006-291x(89)90029-6
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发表时间：
1989
影响因子：
3.1
通讯作者：
Fregien,N
中科院分区：
文献类型：
--
作者：
Ripka,J;Pierce,M;Fregien,N
摘要N-乙酰氨基葡萄糖基转移酶I（GlcNAc-TI）催化细胞表面连接的ASN寡糖从甘露糖终止结构转化为更复杂的碳水化合物结构的第一个反应。突变的中国仓鼠卵巢（CHO）细胞系Lec 1缺乏这种酶，因此，对凝集素伴刀豆球蛋白A的敏感性增加，伴刀豆球蛋白A与Lec 1细胞表面糖蛋白上积累的甘露糖终止寡糖结合。从未观察到Lec 1表型的自发回复突变体。我们在这里报告的分离稳定的回复突变体的Lec 1细胞的亲本CHO细胞凝集素抗性表型后，DNA介导的转化与人类DNA。初级和次级转化体都表达不同水平的GlcNAc-TI酶活性，即使在非选择性条件下培养细胞时，该酶活性也是稳定的。人alu重复DNA序列存在于初级转化体中，但在次级转化体中不能检测到这些序列。
Abstract N-acetylglucosaminyltransferase I (GlcNAc-TI) catalyzes the first reaction in the conversion of ASN-linked cell surface oligosaccharides from a mannose-terminating structure to more complex carbohydrate structures. The mutant Chinese hamster ovary (CHO) cell line, Lec1, is deficient in this enzyme and, therefore, shows increased sensitivity to the lectin, Concanavalin A, which binds to the mannose-terminating oligosaccharides that accumulate on Lec1 cell surface glycoproteins. Spontaneous revertants of the Lec1 phenotype have never been observed. We report here the isolation of stable revertants of Lec1 cells to the parental CHO cell lectin-resistance phenotype after DNA-mediated transformation with human DNA. Both primary and secondary transformants express varying levels of GlcNAc-TI enzyme activity which was stable even when the cells were cultured in nonselective conditions. Human alu repeat DNA sequences are present in the primary transformants, but these sequences could not be detected in the secondary transformants.