Expression of active hBMP2 in transgenic tobacco plants.
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.
Suo
Guangli
G
Center of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100080, China.
Chen
Bing
B
Zhang
Jingyu
J
Gao
Yuan
Y
Wang
Xia
X
He
Zhengquan
Z
Dai
Jianwu
J
D000818:Animals; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018744:DNA, Plant / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors; D018745:Genome, Plant / Q000235:genetics; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D051379:Mice; D030821:Plants, Genetically Modified; D018749:RNA, Plant / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D018744:DNA, Plant; D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; D016212:Transforming Growth Factor beta; D005966:Glucuronidase