Expression of full-length bioactive antimicrobial human lactoferrin in potato plants.
A cDNA fragment encoding human lactoferrin (hLF) linked to a plant microsomal retention signal peptide (SEK-DEL) was stably integrated into the Solanum tuberosum genome by Agrobacterium tumefaciens-mediated leaf disk transformation methods. The lactoferrin gene was expressed under control of both the auxin-inducible manopine synthase (mas) P2 promoter and the cauliflower mosaic virus (CaMV) 35S tandem promoter. The presence of the hLF cDNA in the genome of regenerated transformed potato plants was detected by polymerase chain reaction amplification methods. Full-length hLF protein was identified by immunoblot analysis in tuber tissue extracts from the transformed plants by immunoblot analysis. The hLF produced in transgenic plant tissues migrated during polyacrylamide gel electrophoresis as a single band with an approximate molecular mass equal to hLF. Auxin activation of the mas P2 promoter increased lactoferrin expression levels in transformed tuber and leaf tissues to approximately 0.1% of total soluble plant protein. Antimicrobial activity against four different human pathogenic bacterial strains was detected in extracts of lactoferrin-containing potato tuber tissues. This is the first report of synthesis of full length, biologically active hLF in edible plants.
Chong
D K
DK
Center for Molecular Biology and Gene Therapy, Loma Linda University, CA 92350, USA.
Langridge
W H
WH
D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000378:metabolism; D019677:Artificial Gene Fusion; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D018506:Gene Expression Regulation, Plant; D006801:Humans; D015151:Immunoblotting; D007781:Lactoferrin / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity; D008826:Microbial Sensitivity Tests; D008895:Milk, Human; D010936:Plant Extracts / Q000633:toxicity; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D012482:Salmonella paratyphi A / Q000187:drug effects / Q000254:growth & development; D011198:Solanum tuberosum / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development; D014170:Transformation, Genetic
D010936:Plant Extracts; D021382:Protein Sorting Signals; C062059:SEKDEL sequence; D007781:Lactoferrin