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首页> 外文期刊>African Journal of Biotechnology >Long-term stability of -galactosidase protein expression in the absence of selective growth conditions in transfected Chinese hamster ovary cell
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Long-term stability of -galactosidase protein expression in the absence of selective growth conditions in transfected Chinese hamster ovary cell

机译：在转染中国仓鼠卵巢细胞中没有选择生长条件的情况下 - 高透明酶蛋白表达的长期稳定性

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In this study, the adequacy of β-galactosidase (β-gal) as marker for models that requires durable and high level gene expression in the absence of selective pressure was investigated. Chinese hamster ovary (CHO) cells were transfected with expression vector?pcDNA4/HisMax-TOPO/lacZ containing?lacZ and zeocin resistant genes. 26?recombinant CHO cell lines were established using lipid cationic transfection (TransFast? Transfection Reagent)?as DNA transfer method. 6 clones were productive in the expression of β-gal when grown in the presence of zeocin as the selective pressure. 2 sub-clones, TF8 (1) and TF9 (7) grown for 11 passages in nonselective medium?which?maintained high levels of protein expression with specific β-gal activity in the absence of zeocin were almost constant at?1.5704 and 4.3017 units β-gal activity/mg protein respectively. Specific growth rate of TF8 (1) and TF9 (7) in the absence of zeocin were approximately 0.638 and 0.656 day?1?respectively. The expression of β-gal does not affect the cell growth and the transfectants were stable population in terms of cell viability. Removal of zeocin from the media increases specific growth rate from a range of 24 to 52% and β-gal protein production in reference to specific activity increases from 128 to 320% with the capability to be expanded to larger volumes. In this study,?we demonstrate transfected CHO cells with the ability to produce?β-gal without the presence of zeocin for at least 11 passages.
机译：在本研究中，研究了在没有选择性压力的情况下需要耐久性和高水平基因表达的模型中β-半乳糖苷酶（β-GAL）作为标记的充分性。用表达载体转染中国仓鼠卵巢（CHO）细胞？PCDNA4 / Hismax-Topo / LacZ含有αSacZ和Zeocin抗性基因。 26？使用脂质阳离子转染（Transfastα转染试剂）建立重组CHO细胞系列？作为DNA转移方法。在Zeocin存在作为选择性压力时，在β-gal的表达中施加6克隆的克隆。在非选择性培养基中生长的2个亚克隆，TF8（1）和TF9（7）增加了11个通道？哪种α保持高水平的蛋白质表达，在没有Zeocin的情况下，在没有Zeocin的情况下与β-1.5704和4.3017单位β-加仑活性/ mg蛋白质。在没有Zeocin的情况下，TF8（1）和TF9（7）的特异性生长速率约为0.638和0.656天？ β-gal的表达不会影响细胞生长，并且在细胞活力方面，转染剂是稳定的群体。从培养基中除去Zeocin从24-52％的范围内增加比例，参考特定活性的β-加仑蛋白质产生从128增加到320％，其能力扩展到较大的体积。在这项研究中，我们证明了转染的CHO细胞，其能够在没有Zeocin的情况下生产α-加仑的能力至少11个通道。

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