泛染色体开放元件对CHO细胞株中抗体表达量的影响
Influence of ubiquitous chromatin opening element on the antibody expression in CHO cells
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摘要: 泛染色体开放元件(UCOE)来自于人类看家基因启动子基因序列，能阻止基因转录沉默，使转录的目的基因不受“位置效应”的影响，从而使目的蛋白在细胞中高表达。本研究考察了不同片段大小的UCOE元件对稳转细胞株中抗体表达量的影响。将UCOE元件中来自于CBX3基因的1.5 kb片段和来自于HNRPA2B1基因的2.5 kb片段及完整UCOE 4.0 kb片段分别插入到含有抗体轻、重链的质粒载体中，转染中国仓鼠卵巢(CHO)细胞，加压筛选获得4组单克隆细胞株。比较不同组的单克隆细胞株抗体表达量，结果显示，转染UCOE 1.5 kb、2.5 kb的单克隆细胞株抗体表达量是对照组的1.5~2倍，转染UCOE 4.0 kb的单克隆细胞株抗体表达量是对照组的3~4倍；两个看家基因启动子基因序列对目的基因表达的增强作用可相互叠加。Abstract: Ubiquitous chromatin opening element(UCOE), composed of the promoters of human housekeeping genes, prevents transgene from silencing and produces consistent, stable and high-level gene expression irrespective of the chromosomal integration site. The research studied the influence of different UCOE element parts on antibody expression in CHO cells. UCOE 1. 5 kb from chromobox homolog 3(CBX3), UCOE 2. 5 kb from the heterogeneous ribonucleoprotein A2/B1(HNRPA2B1)and the whole UCOE 4. 0 kb were inserted into the antibody light and heavy chain vectors, respectively, and transfected into CHO cells using antibiotics-Zeocin and Blasticidin for pressure selection. Four groups of monoclonal cells were harvested and antibody expression of each group was detected. The monoclonal cells with UCOE 1. 5 kb and UCOE 2. 5 kb increased 1. 5 to 2-fold in the level of antibody expression, wheareas, monoclonal cells with UCOE 4. 0 kb increased 3 to 4-fold. The enhancement of two housekeeping promoter genes on antibody expression could stack up.
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