重组人白细胞介素-7在中国仓鼠卵巢（CHO）细胞中的克隆与表达
Cloning and Expression of Recombinant Human Interleukin-7 in Chinese Hamster Ovary (CHO) Cells.
作者信息
Toghraie Fatemeh Sadat, Sharifzadeh Seyedeh Masoumeh, Ramezani Amin, Mahmoudi Maymand Elham, Yazdanpanah-Samani Mahsa, Ghaderi Abbas
机构信息
Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical science, Shiraz, Iran.
Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
出版信息
Rep Biochem Mol Biol. 2017 Oct;6(1):66-73.
BACKGROUND
The critical role of interleukin-7 (IL-7) in homeostatic proliferation and T cell survival has made it a promising cytokine for the treatment of various clinical conditions, especially those associated with lymphopenia.
METHODS
In the present study we expressed recombinant human interleukin-7 (rhIL-7) in Chinese hamster ovary (CHO)-K1 cells. CHO-K1 cells were stably transfected with both circular and linear forms of the pBud-h-7 recombinant by electroporation. Expression of rhIL-7 in CHO-K1 cells was confirmed by enzyme-linked immunosorbent assay (ELISA) and dot and western blots.
RESULTS
On western blots of transformed cells, a single 25 kDa band was observed, consistent with the expected molecular weight of glycosylated hIL-7. No significant expression difference was observed between cells transfected with circular or linear plasmids.
CONCLUSION
We established a stable CHO-K1 cell line expressing rhIL-7, which we consider to be a promising system for the production of rhIL-7 as a biopharmaceutical.
背景
白细胞介素-7（IL-7）在稳态增殖和T细胞存活中起关键作用，这使其成为治疗各种临床病症，尤其是与淋巴细胞减少相关病症的一种有前景的细胞因子。
方法
在本研究中，我们在中国仓鼠卵巢（CHO）-K1细胞中表达重组人白细胞介素-7（rhIL-7）。通过电穿孔将pBud-h-7重组体的环状和线性形式稳定转染至CHO-K1细胞。通过酶联免疫吸附测定（ELISA）以及斑点印迹和蛋白质印迹证实rhIL-7在CHO-K1细胞中的表达。
结果
在转化细胞的蛋白质印迹上，观察到一条单一的25 kDa条带，与糖基化hIL-7的预期分子量一致。用环状或线性质粒转染的细胞之间未观察到明显的表达差异。
结论
我们建立了一种表达rhIL-7的稳定CHO-K1细胞系，我们认为这是生产作为生物药物的rhIL-7的一种有前景的系统。
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