Expression and localization of human lysozyme in the endosperm of transgenic rice.
In order to understand the characteristics of recombinant protein expression and sublocalization in rice ( Oryza sativa L.) endosperm, we examined the expression level of human lysozyme protein and its subcellular location in transgenic rice seeds driven by rice glutelin and globulin promoters and signal peptides. A time course of human lysozyme expression during endosperm development was analyzed. The results showed that the expression profile of recombinant protein accumulation in endosperm paralleled that of the two storage proteins. Immunofluorescence microscopy revealed that human lysozyme and storage proteins co-localized to type-II protein bodies. Both promoter-signal peptide parings targeted recombinant protein to the protein bodies. In addition, a transgenic line with a higher lysozyme expression level exhibited morphologically different protein bodies with an unbalanced composition of lysozyme and native storage proteins. The high-level expression of recombinant protein distorted the trafficking and sorting of native storage proteins in rice endosperm and affected the expression of native storage protein.
Yang
Daichang
D
Ventria Bioscience, 4110 North Freeway, Sacramento, CA 95834, USA. dyang@ventriabio.com
Guo
Fengli
F
Liu
Bo
B
Huang
Ning
N
Watkins
Simon C
SC
D015971:Gene Expression Regulation, Enzymologic; D005916:Globulins / Q000235:genetics; D005983:Glutens / Q000235:genetics; D006801:Humans; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009113:Muramidase / Q000235:genetics* / Q000378:metabolism; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure
D005916:Globulins; D010940:Plant Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D005983:Glutens; D009113:Muramidase