Production of recombinant human lactoferin in transgenic barley

Abstract

Introduction of human lactoferrin (hLF) gene, a cationic iron-binding glycoprotein, in the genome of commercial Bulgarian barley (Hordeum vurgare L.) cultivar was performed via particle bombardment. A rapid, efficient, and reproducible system was used to generate green, fertile, transgenic plants. Mature zygotic embryos from dry seeds of malting cultivar Ruen were bombarded with two plasmids (pAFT105 and pUBLF) containing human lactoferrin gene. The embryos were co-transformed either with pDM302 containing selectable marker gene (bar) for herbicide resistance and plasmid pAct1-F carrying uidA gene forβ-glucuronidase activity, or with plasmid pAHC25 including bar and uidA genes both under the control of the ubiquitin (Ubi) promoter. Forty eight individual regenerants were selected on Bialaphos (glufosinate-ammonium) and subjected to PCR analysis using primers corresponding to the sequence of hLF gene. The introgression of hLF cDNA was confirmed in 27 plants from 16 lines. The expression of recombinant human lactoferrin was proved by Southern and Western blot analyses. Transmission of the hLF gene to T1 progeny was studied. In this study we present an efficient system for introduction of gene encoding valuable recombinant protein. These data suggest that commercial barley can be used as a bioreactor for molecular farming ofpharmaceutical proteins. © 2007 Taylor and Francis Group, LLC.