Zhang Lijuan, Lawrence Roger, Frazier Beth A, Esko Jeffrey D
Washington University School of Medicine, St. Louis, MO, USA.
Methods Enzymol. 2006;416:205-21. doi: 10.1016/S0076-6879(06)16013-9.
Most glycosaminoglycan (GAG)-defective mutants have been isolated and characterized from Chinese hamster ovary (CHO) cells. Wild-type and GAG-defective CHO cells have been used by several hundreds of laboratories to study how altering the GAG structure of proteoglycans affects fundamental properties of cells, such as bacterial/viral infection, signaling, protein degradation, and cell adhesion. This chapter describes methods used to construct and characterize new CHO cell lines with gain-of-function GAG structures. These novel CHO cell lines allow herpes simplex virus (HSV) entry or have anticoagulant properties that are not possessed by wild-type CHO cells. The method used to study GAG biosynthetic mechanisms that control specific GAG sequence assembly is also described.
大多数糖胺聚糖（GAG）缺陷型突变体是从中国仓鼠卵巢（CHO）细胞中分离和鉴定出来的。数百个实验室使用野生型和GAG缺陷型CHO细胞来研究改变蛋白聚糖的GAG结构如何影响细胞的基本特性，如细菌/病毒感染、信号传导、蛋白质降解和细胞粘附。本章描述了构建和鉴定具有功能获得性GAG结构的新型CHO细胞系的方法。这些新型CHO细胞系允许单纯疱疹病毒（HSV）进入，或具有野生型CHO细胞所不具备的抗凝血特性。还描述了用于研究控制特定GAG序列组装的GAG生物合成机制的方法。
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