骨形态发生蛋白10及其突变体在CHO细胞中的表达

Expression of Bone Morphogenetic Protein 10 and Its Mutants in CHO Cells

摘要:

为了解决骨形态发生蛋白10（bone morphogenetic protein 10，BMP10）前体蛋白proBMP10在中国仓鼠卵巢（CHO）细胞中由于Furin导致的表达产物不均一问题，构建了proBMP10 在Furin酶切识别位点处的突变体proBMP10-1（R313K）和proBMP10-2（R316K）,并分别将目的基因定点整合进CHO-K1-BAK^-/BAX^-基因组，成功构建了稳定表达目标蛋白质的重组CHO细胞株。结果表明，proBMP10-2（R316K）不再被Furin切割，且具有生物活性，而proBMP10-1（R313K）仍然会被Furin切割。

Abstract:

To address the issue of heterogeneous expression products derived from proBMP10 (the precursor protein of bone morphogenetic protein 10, BMP10) in Chinese hamster ovary(CHO) cells caused by Furin, the mutations of proBMP10 at its Furin recognition sites, proBMP10-1 (R313K) and proBMP10-2(R316K), were constructed and site-directedly integrated into the genome of CHO-K1-BAK^-/BAX^- respectively. The recombinant CHO cell lines stably expressing the target proteins were successfully established. The results showed that the proBMP10-2 (R316K) was no longer cleaved by Furin and retained biological activity, while proBMP10-1 (R313K) continued to be cleaved by Furin.