Abstract
Heterologous protein expression levels in transgenic plants are of critical importance in the production of plant-made pharmaceuticals (PMPs). We studied a puroindoline b promoter and signal peptide (Tapur) driving human lysozyme expression in rice endosperm. The results demonstrated that human lysozyme expressed under the control of the Tapur cassette is seed-specific, readily extractable, active, and properly processed. Immuno-electron microscopy indicated that lysozyme expressed from this cassette is localized in protein bodies I and II in rice endosperm cells, demonstrating that this non-storage promoter and signal peptide can be used for targeting human lysozyme to rice protein bodies. We successfully employed a strategy to improve the expression of human lysozyme in transgenic rice grain by combining the Tapur cassette with our well established Gt1 expression system. The results demonstrated that when the two expression cassettes were combined, the expression level of human lysozyme increased from 5.24 ± 0.34 mg−1 g flour for the best single cassette line to 9.24 ± 0.06 mg−1 g flour in the best double cassette line, indicating an additive effect on expression of human lysozyme in rice grain.
Similar content being viewed by others
References
RK Chikwamba MP Scott LB Mejia HS Mason K Wang (2003) ArticleTitleLocalization of a bacterial protein in starch granules of transgenic maize kernels Proc Natl Acad Sci USA 100 11127–11132 Occurrence Handle10.1073/pnas.1836901100 Occurrence Handle12960364
DK Chong WH Langridge (2000) ArticleTitleExpression of full-length bioactive antimicrobial human lactoferrin in potato plants Transgenic Res 9 71–78 Occurrence Handle10.1023/A:1008977630179 Occurrence Handle10853271
JF Digeon E Guiderdoni R Alary N Michaux-Ferriere P Joudrier MF Gautier (1999) ArticleTitleCloning of a wheat puroindoline gene promoter by IPCR and analysis of promoter regions required for tissue-specific expression in transgenic rice seeds Plant Mol Biol 39 1101–1112 Occurrence Handle10.1023/A:1006194326804 Occurrence Handle10380798
S Dellaporta J Wood J Hick (1983) ArticleTitleA plant DNA mini preparation: version II Plant Mol Biol Report 1 19–21
L Dubreil T Gaborit B Bouchet DJ Gallant WF Broekaert L. Quillien et al. (1998) ArticleTitleSpatial and temporal distribution of the major isoforms of puroindolines (puroindoline-a and puroindoline-b) and non specific lipid transfer protein (ns-LTPle(1)) of Triticum aestivum seeds. Relationships with their in vitro antifungal properties Plant Sci 138 121–135 Occurrence Handle10.1016/S0168-9452(98)00121-6
EE Hood DR Witcher S Maddock T Meyer C Baszczynski M. Bailey et al. (1997) ArticleTitleCommercial production of avidin from transgenic maize: characterization of transformant, production, processing, extraction and purification Mol Breed 3 291–306 Occurrence Handle10.1023/A:1009676322162
H Horvath J Huang O Wong E Kohl TW Okita CG. Kannangara et al. (2000) ArticleTitleThe production of recombinant proteins in transgenic barley grains Proc Natl Acad Sci USA 97 1914–1919 Occurrence Handle10.1073/pnas.030527497 Occurrence Handle10677555
J Huang S Nandi L Wu D Yalda G Bartley RL. Rodriguez et al. (2002) ArticleTitleExpression of natural antimicrobial human lysozyme in rice grain Mol Breed 10 83–94 Occurrence Handle10.1023/A:1020355511981
N Huang L Wu S Nandi E Bowman J Huang T. Sutliff et al. (2001) ArticleTitleThe tissue-specific activity of a rice beta-glucanase promoter (Gns9) is used to select rice transformants Plant Sci 161 589–595 Occurrence Handle10.1016/S0168-9452(01)00447-2
Y-S Hwang D Yang C McCullar L Wu L Chen P. Pham et al. (2002) ArticleTitleAnalysis of the rice endosperm-specific globulin promoter in transformed rice cells Plant Cell Report 20 842–847 Occurrence Handle10.1007/s00299-001-0406-z
BO Juliano (1985) Polysaccharides, Proteins and Lipids of Rice American Association of Cereal Chemistry St. Paul, MN
K Krishnamurthy C Balconi C Balconi JE Sherwood MJ Giroux (2001) ArticleTitleWheat puroindolines enhance fungal disease resistance in transgenic rice Mol Plant-Microbe Interact 14 1255–1260 Occurrence Handle11605965
K Krishnamurthy MJ Giroux (2001) ArticleTitleExpression of wheat puroindoline genes in transgenic rice enhances grain softness Nat Biotechnol 19 162–166 Occurrence Handle10.1038/84435 Occurrence Handle11175732
A Leite EL Kemper Silva MJ da AD Luchessi RMP Siloto ED. Bonaccorsi et al. (2000) ArticleTitleExpression of correctly processed human growth hormone in seeds of transgenic tobacco plants Mol Breed 6 47–53 Occurrence Handle10.1023/A:1009648532210
XX Li VR Franceschi TW Okita (1993) ArticleTitleSegregation of storage protein messenger RNAs on the rough endoplasmic reticulum membranes of rice endosperm cells Cell 72 869–879 Occurrence Handle10.1016/0092-8674(93)90576-C Occurrence Handle8458081
R Matsushima M Kondo M Nishimura L Hara-Nishimura (2003) ArticleTitleA novel ER-derived compartment, the ER body, selectively accumulates a β-glucosidase with an ER-retention signal in Arabidopsis Plant J 33 493–503 Occurrence Handle10.1046/j.1365-313X.2003.01636.x Occurrence Handle12581307
J Messeguer C Fogher E Guiderdoni V Marfa MM Catala G. Baldi et al. (2001) ArticleTitleField assessments of gene flow from transgenic to cultivated rice (Oryza sativa L.) using a herbicide resistance gene as tracer marker Theor Appl Genet 103 1151–1159 Occurrence Handle10.1007/s001220100713
K Müntz (1998) ArticleTitleDeposition of storage proteins Plant Mol Biol 38(1–2) 77–99 Occurrence Handle10.1023/A:1006020208380
S Nandi Y Suzuki J Huang D Yalda P Pham L. Wu et al. (2002) ArticleTitleExpression of human lactoferrin in transgenic rice grains for application in infant formula Plant Sci 163 713–722 Occurrence Handle10.1016/S0168-9452(02)00165-6
J-M Neuhaus JC Rogers (1998) ArticleTitleSorting of proteins to vacuoles in plant cells Plant Mol Biol 38 127–144 Occurrence Handle10.1023/A:1006032627036 Occurrence Handle9738964
A Ritala AM Nuutila R Aikasalo V Kauppinen J Tammisola (2002) ArticleTitleMeasuring gene flow in the cultivation of transgneic barley Crop Sci 42 278–285 Occurrence Handle11756286
S Romer PD Fraser JW Kiano CA Shipton N Misawa W. Schuch et al. (2000) ArticleTitleElevation of the provitamin A content of transgenic tomato plants Nature Biotechnol 18 666–669 Occurrence Handle10.1038/76523
PR Shewry JA Napier AS Tatham (1995) ArticleTitleSeed Storage Proteins–Structures and Biosynthesis Plant Cell 7 945–956 Occurrence Handle10.1105/tpc.7.7.945 Occurrence Handle7640527
D Shugar (1952) ArticleTitleThe measurement of lysozyme activity and the ultra-violet inactivation of lysozyme Biochim Biophys Acta 8 302–309 Occurrence Handle10.1016/0006-3002(52)90045-0 Occurrence Handle14934741
E Stoger C Vaquero E Torres M Sack L Nicholson J. Drossard et al. (2000) ArticleTitleCereal crops as viable production and storage systems for pharmaceutical scFv antibodies Plant Mol Biol 42 583–590 Occurrence Handle10.1023/A:1006301519427 Occurrence Handle10809004
Y Tada S Utsumi F Takaiwa (2003) ArticleTitleForeign gene products can be enhanced by introduction into low storage protein mutants Plant Biotechnol 1 411–422 Occurrence Handle10.1046/j.1467-7652.2003.00038.x
A Vitale NV Raikhel (1999) ArticleTitleWhat do proteins need to reach different vacuoles Trends Plant Sci 4 IssueID4 149–155 Occurrence Handle10.1016/S1360-1385(99)01389-8 Occurrence Handle10322549
D Wettstein Particlevon G Mikhaylenko JA Froseth C Kannangar (2000) ArticleTitleImproved barley broiler feed with transgenic malt containing heat-stable (1.3–1.4)-β-glucanase Proc Natl Acad Sci USA 97 13511–13517
D Yang F Guo B Liu N Huang SC Watkins (2003) ArticleTitleExpression and localization of human lysozyme in the endosperm of transgenic rice Planta 216 597–603 Occurrence Handle12569401
D Yang L Wu YS Hwang L Chen N Huang (2001) ArticleTitleExpression of the REB transcriptional activator in rice grains improves the yield of recombinant proteins whose genes are controlled by a Reb responsive promoter Proc Natl Acad Sci USA 98 11438–11443 Occurrence Handle10.1073/pnas.201411298 Occurrence Handle11572990
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hennegan, K., Yang, D., Nguyen, D. et al. Improvement of Human lysozyme Expression in Transgenic Rice Grain by Combining Wheat (Triticum aestivum) puroindoline b and Rice (Oryza sativa) Gt1 Promoters and Signal Peptides. Transgenic Res 14, 583–592 (2005). https://doi.org/10.1007/s11248-004-6702-y
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11248-004-6702-y