Expression and production of recombinant human interleukin-2 in potato plants.
Interleukin-2 is a pharmacologically important cytokine secreted by T lymphocytes. Recombinant interleukin-2 has been produced and found to be useful for many medical applications. Mass production of recombinant interleukin-2 will be prerequisite for a wider application of this molecule. In this study we investigated the possibility of using potato tubers for the production of recombinant human interleukin-2 in large quantity. A binary vector carrying the human interleukin-2 gene under a potato tuber-specific promoter (patatin promoter) was constructed. Several potato transformants expressing the human interleukin-2 gene were generated by Agrobacterium-mediated transformation. Expression of the human interleukin-2 gene was confirmed by Northern blotting and the protein level was determined by Western blot analyses. A bioassay revealed that human interleukin-2 expressed in the potato tuber supported proliferation of interleukin-2-dependent cells, CTLL-2. We found that the recombinant protein in the 2-week-old microtuber has the highest activity (115 units per gram of microtuber) and estimated that an average yield for a potato (average 200 g per potato) was 23,000 units of rhIL-2 activity. The results suggest that the potato tuber is an excellent system for the mass production of biologically active human interleukin-2.
Park
Yoonkyung
Y
Department of Genetic Engineering, Chosun University, Kwang-Ju, Korea.
Cheong
Hyeonsook
H
D001681:Biological Assay; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D011198:Solanum tuberosum; D013997:Time Factors
D007376:Interleukin-2; D011994:Recombinant Proteins; D012313:RNA