Haploid technology allows for the efficient and rapid generation of homozygous antibody-accumulating transgenic tobacco plants.
The large-scale production of plant-derived recombinant proteins requires the breeding of lines homozygous for the transgene(s). These can be selected by progeny testing over multiple sexual generations, but a more efficient means is to fix homozygosity in a single generation using doubled haploid technology. In this study, transgenic tobacco plants, hemizygous for both of the independently inherited genes encoding the light and heavy chains of the anti-human immunodeficiency virus monoclonal antibody 2F5, were used to establish embryogenic pollen cultures. The improved protocol employed in this study guaranteed a very high regeneration efficiency, with more than 50% of the regenerants being spontaneously doubled haploids. Hence, there was no requirement to chemically induce chromosome doubling to recover sufficient entirely homozygous recombinants. As expected, approximately 25% of the regenerants were homozygous for both transgenes. Thus, the employment of haploid technology allowed for the efficient and rapid generation of true-breeding tobacco lines accumulating functional immunoglobulins.
Floss
Doreen M
DM
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Phytoantibodies, Gatersleben, Germany.
Kumlehn
Jochen
J
Conrad
Udo
U
Saalbach
Isolde
I
D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant; D006238:Haploidy; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011058:Pollen / Q000166:cytology / Q000196:embryology; D016133:Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
D000906:Antibodies