title: Molecular cloning and transgenic expression of a synthetic human erythropoietin gene in tobacco.
abstract: Erythropoietin (EPO) is a hormone belonging to a group of hematopoietic growth factors that control the proliferation and differentiation of bone marrow cells. It induces the production of erythrocytes, thereby increasing the amount of circulating hemoglobin and oxygen. Previous attempts to transgenically express human EPO in plants failed to succeed because the plants exhibited abnormal morphology and infertility. In the present work, we describe the generation of fertile transgenic tobacco plants able to express a synthetic version of human EPO. A 582-bp fragment of the human EPO gene was synthesized using a PCR-based method and ligated into pCR-Blunt. After sequencing, the human EPO fragment was transferred to pWUbi.tm1 and the expression cassette was then transferred to the binary vector pWBVec4a. After Agrobacterium-mediated transformation of Nicotiana tabacum SR1 plants, integration of the transgene into T(0) and T(1) plant genomes was confirmed by PCR. The human EPO gene was found to be expressed in tobacco leaves at the mRNA and protein levels. Self-crossing allowed us to obtain T(1) plants exhibiting Mendelian segregation of the transgene. None of the plants presented any kind of malformation or deformity.
authors: Graduate Program in Cell and Molecular Biology, Biotechnology Center, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil. fesperb@ig.com.br
authors: Fernanda
authors: F
authors: Sperb
authors: Isabel C R
authors: IC
authors: Werlang
authors: Marcia
authors: M
authors: Margis-Pinheiro
authors: Luiz A
authors: LA
authors: Basso
authors: Diógenes S
authors: DS
authors: Santos
authors: Giancarlo
authors: G
authors: Pasquali