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

共 38 条

[21] Simple, rapid, high-purity preparation of recombinant human platelet-derived growth factor-BB [J].

Karumuri, Nagaraju N. ;

Gangireddy, Srinivasa R. ;

Narala, Venkata R. ;

Majee, Sangita S. ;

Gunwar, Sripad ;

Reddy, Raju C. .

BIOTECHNOLOGY LETTERS, 2007, 29 (09) :1333-1339

[22] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[23] Platelet derived growth factor receptor alpha is essential for establishing a microenvironment that supports definitive erythropoiesis [J].

Li, Wen-Ling ;

Yamada, Yoshihiro ;

Ueno, Masaya ;

Nishikawa, Satomi ;

Nishikawa, Shin-Ichi ;

Takakura, Nobuyuki .

JOURNAL OF BIOCHEMISTRY, 2006, 140 (02) :267-273

[24]

Mandracchia V J, 2001, Clin Podiatr Med Surg, V18, P189

[25] Agrobacterium-mediated transformation as a tool for functional genomics in fungi [J].

Michielse, CB ;

Hooykaas, PJJ ;

van den Hondel, CAMJJ ;

Ram, AFJ .

CURRENT GENETICS, 2005, 48 (01) :1-17

[26]

Nevalainen KMH, 2005, TRENDS BIOTECHNOL, V23, P468, DOI 10.1016/j.tibtech.2005.06.002

[27]

OSTMAN A, 1988, J BIOL CHEM, V263, P16202

[28] Activators and target genes of Rel/NF-κB transcription factors [J].

Pahl, HL .

ONCOGENE, 1999, 18 (49) :6853-6866

[29]

Peng F., 2008, EUROPEAN J NEUROSCIE, V7, P1255

[30] Filamentous fungi as cell factories for heterologous protein production [J].

Punt, PJ ;

van Biezen, N ;

Conesa, A ;

Albers, A ;

Mangnus, J ;

van den Hondel, C .

TRENDS IN BIOTECHNOLOGY, 2002, 20 (05) :200-206

← 1 2 3 4 →