Expression and Production of Therapeutic Recombinant Human Platelet-Derived Growth Factor-BB in Pleurotus eryngii

被引:5
作者
Choi, Jun-Hui [1 ]
Kim, Seung [2 ]
Sapkota, Kumar [1 ,3 ]
Park, Se-Eun [1 ]
Kim, Sung-Jun [1 ]
机构
[1] Chosun Univ, Dept Biotechnol, Kwangju 501759, South Korea
[2] Gwangju Univ, Dept Alternat Med, Kwangju 503703, South Korea
[3] Tribhuvan Univ, Cent Dept Zool, Kathmandu, Nepal
关键词
Agrobacterium tumefaciens; Human platelet-derived growth factor-BB; Pleurotus eryngii; Gene transfer; Gene expression; MEDIATED TRANSFORMATION; TARGET;
D O I
10.1007/s12010-011-9279-y
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Recombinant human platelet-derived growth factor-BB (rhPDGF-BB) is widely used in many therapeutic applications. Until now, there has been no report on rhPDGF-BB expressed in fungi. In this study, we tested whether Pleurotus eryngii could support the expression of human therapeutic rhPDGF-BB protein. A binary vector pCAMBIA1304 containing the hPDGF-BB gene was constructed and introduced into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformation of hPDGF-BB gene was confirmed by Southern blot and PCR, whereas the expression was confirmed by Western blot analysis. The recombinant hPDGF-BB reached a maximum expression level of 1.98% of total soluble protein in transgenic mycelia and was in dimeric form. A bioassay revealed that hPDGF-BB expressed in P. eryngii increased proliferation of NIH-3T3 cells similarly to standard material. These results suggest that P. eryngii can be a robust system for the production of human therapeutic proteins including the hPDGF-BB.
引用
收藏
页码:611 / 623
页数:13
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