Expression of Bone Morphogenetic Protein 10 and Its Mutants in CHO Cells
段珂 1段作营 1金坚2
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作者信息
1. 江南大学生物工程学院,江苏无锡 214122
2. 江南大学生命科学与健康工程学院,江苏无锡 214122
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摘要
为了解决骨形态发生蛋白10(bone morphogenetic protein 10,BMP10)前体蛋白proBMP 10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 proBMP 10(the precursor protein of bone morphogenetic protein 10,BMP 10)in Chinese hamster ovary(CHO)cells caused by Furin,the mutations of proBMP 10 at its Furin recognition sites,proBMP 10-1(R313K)and proBMP 10-2(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 proBMP 10-2(R316K)was no longer cleaved by Furin and retained biological activity,while proBMP 10-1(R313K)continued to be cleaved by Furin.
关键词
骨形态发生蛋白10/突变体/定点整合
Key words
bone morphogenetic protein 10/mutant/site-specific integration
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