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

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2011年 / 165卷 / 02期

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

页码：611 / 623

页数：13

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