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Expression of active hBMP2 in transgenic tobacco plants

  • Genetic transformation and Hybridization
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Abstract

Bone morphogenetic protein 2 (BMP2) is important for bone tissue repair. The goal of this research is to construct a high level human BMP2 (hBMP2) expression system using transgenic tobacco plants as a bioreactor. Cauliflower mosaic virus (CaMV) 35S promoter, alfalfa mosaic virus (AMV) enhancer, tobacco mosaic virus (TMV) enhancer, matrix attachment regions (MARs) sequence, and “Kozak” sequence were used to construct recombinant expression vectors and the high-expression vectors were screened out through GUS-fusions assay. The promoter is the most important factor; double-CaMV 35S promoter is more effective than single promoter. The AMV or TMV enhancer is able to promote the foreign protein expression. After four-step purification, the activated hBMP2 (0.02% total soluble protein) was obtained. Our results suggested that the transgenic tobacco has great potential to be used as a bioreactor to produce hBMP2.

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Abbreviations

kDa:

kilodalton(s)

BMP2:

Bone morphogenetic protein-2

CUTG:

Codon Usage Tabulated from GenBank

CAI:

Codon adaptation index

GUS :

β-Glucuronidase

CaMV35S promoter:

Cauliflower mosaic virus 35S promoter

AMV:

Alfalfa mosaic virus

TMV:

Tobacco mosaic virus

MU:

Metylumbelliferone

EK:

Enterokinase

h :

Human

M :

Modified

N :

Native

Sarcosyl:

Sodium lauroyl sarcosine

ALP:

Alkaline phosphatase

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Acknowledgements

This work was supported by Chinese Academy of Sciences’ “100 talented Scholar Program” and Chinese Academy of Sciences’ “Knowledge Innovation Program” Grants (No. KSCX2-SW-205; KSCW2-SW-218), and also Supported by NSFC (30428017), and by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry. The authors also gratefully acknowledge the support of K. C. Wong Education Foundation, Hong Kong

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Authors and Affiliations

  1. Center of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100080, China

    Guangli Suo, Bing Chen, Jingyu Zhang, Yuan Gao, Xia Wang & Jianwu Dai

  2. Biotechnology Research Center, China Three Gorges University, Yichang, 443002, China

    Zhengquan He

  3. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 3 Nanyitiao, Zhongguancun, Beijing, 100080, China

    Jianwu Dai

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  1. Guangli Suo
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  2. Bing Chen
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  3. Jingyu Zhang
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  4. Yuan Gao
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  5. Xia Wang
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  6. Zhengquan He
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  7. Jianwu Dai
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Correspondence to Jianwu Dai.

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Communicated by F. Sato

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Suo, G., Chen, B., Zhang, J. et al. Expression of active hBMP2 in transgenic tobacco plants. Plant Cell Rep 25, 1316–1324 (2006). https://doi.org/10.1007/s00299-006-0173-y

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  • DOI: https://doi.org/10.1007/s00299-006-0173-y

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