Synthesis and expression of the gene for human epidermal growth factor in transgenic potato plants

Salmanian, Ali-Hatef; Guschin, Alexander; Medvedeva, T.; Noori–Daloii, Mohammad Reza; Domansky, N. N.

doi:10.1007/bf00129738

Cited by 6 publications

(2 citation statements)

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“…The successful expression of EGF has been reported in tobacco ( Nicotiana tabacum and N. benthamiana ) [ 9 , 10 , 15 , 19 , 45 , 46 ], tomato [ 12 ], soybean [ 16 ], potato [ 47 ], rice [ 20 , 48 ], and saffron [ 21 ]. The strategies for the expression of EGF in plants were either transient or stable.…”

Section: Introductionmentioning

confidence: 99%

“…The transient methods reported used either plant-virus-based vectors [ 9 , 46 ] or mini-Ti vectors [ 15 ]. The stable expression of EGF in plants adopted both nuclear [ 9 , 11 , 12 , 13 , 16 , 20 , 21 , 45 , 46 , 47 , 48 ] and chloroplast [ 10 , 19 , 49 ] transformation. The expression levels varied from 0.001% to 7.8% of total soluble proteins (TSP).…”

Section: Introductionmentioning

confidence: 99%

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Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts

et al. 2023

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Background Chloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplast transformation approach at high expression levels. However, high expression of epidermal growth factor (EGF) in chloroplasts with the technology is still unavailable. Results The present work explored the high-level expression of recombinant EGF, a protein widely applied in many clinical therapies, in tobacco chloroplasts. In this work, homoplastic transgenic plants expressing fusion protein GFP-EGF, which was composed of GFP and EGF via a linker, were generated. The expression of GFP-EGF was confirmed by the combination of green fluorescent observation and Western blotting. The achieved accumulation of the recombinant fusion GFP-EGF was 10.21 ± 0.27% of total soluble proteins (1.57 ± 0.05 g kg− 1 of fresh leaf). The chloroplast-derived GFP-EGF was capable of increasing the cell viability of the NSLC cell line A549 and enhancing the phosphorylation level of the EGF receptor in the A549 cells. Conclusion The expression of recombinant EGF in tobacco chloroplasts via chloroplast transformation method was achieved at considerable accumulation level. The attempt gives a good example for the application of chloroplast transformation technology in recombinant pharmaceutical protein production.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts

et al. 2023

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Improved expression of recombinant plant-made hEGF

Thomas

Walmsley

2014

Plant Cell Rep

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The yield of recombinant hEGF was increased approximately tenfold through a range of optimisations. Further, the recombinant protein was found to have biological activity comparable to commercial hEGF. Human epidermal growth factor (hEGF) is a powerful mitogen that can enhance the healing of a wide range of injuries, including burns, cuts, diabetic ulcers and gastric ulcers. However, despite its clinical value, hEGF is only consistently used for the treatment of chronic diabetic ulcers due to its high cost. In this study, hEGF was transiently expressed in Nicotiana benthamiana plants and targeted to the apoplast, ER and vacuole. Several other approaches were also included in a stepwise fashion to identify the optimal conditions for the expression of recombinant hEGF. Expression was found to be highest in the vacuole, while targeting hEGF to the ER caused a decrease in total soluble protein (TSP). Using a codon optimised sequence was found to increase vacuolar targeted hEGF yield by ~34 %, while it was unable to increase the yield of ER targeted hEGF. The use of the P19 silencing inhibitor was able to further increase expression by over threefold, and using 5-week-old plants significantly increased expression compared to 4- or 6-week-old-plants. The combined effect of these optimisations increased expression tenfold over the initial apoplast targeted construct to an average yield of 6.24 % of TSP. The plant-made hEGF was then shown to be equivalent to commercial E. coli derived hEGF in its ability to promote the proliferation of mouse keratinocytes. This study supports the potential for plants to be used for the commercial production of hEGF, and identifies a potential limitation for the further improvement of recombinant protein yields.

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Humanizing infant milk formula to decrease postnatal HIV transmission

Blais

Altosaar

2007

Trends in Biotechnology

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Synthesis and expression of the gene for human epidermal growth factor in transgenic potato plants

Cited by 6 publications

References 13 publications

Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts

Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts

Improved expression of recombinant plant-made hEGF

Humanizing infant milk formula to decrease postnatal HIV transmission

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