Expression of Human Glucagon-like Peptide-1 in Transgenic Tobaccos

Abstract

Abstract: Human pancreatic glucagon-like peptide-1(hGLP1)is a short peptide hormone with special attention in the treatmentof diabetes, is considered as the most promising candidate medicines. Based on design composition of hGLP1 gene and construction of plantexpression vector, hGLP1 gene was introduced into tobacco through Agrobacterium tumfaciens-mediated method. The PCR amplificationand Southern blot analysis indicated that the hGLP1 gene have been integrated into the genome of six lines of the regenerated tobaccos. GUShistochemistry staining showed that the reporter gene fusion with the target gene was expressed in transgenic tobacco plants. Western blotanalysis confirmed that hGLP1 fusion protein was detected in two lines of transgenic tobacco leaves. The fusion protein demonstrated a certainbiological activity in reducing blood sugar by elementary animal experiments.

Key words: Tobacco , Transgene , Human glucagon-like peptide 1 , Biological activity

Liang Hongwei Chen Faju Wang Yubing Li Fenglan, Lu Hai. Expression of Human Glucagon-like Peptide-1 in Transgenic Tobaccos[J]. Biotechnology Bulletin, 2013, 0(1): 139-143.

References

[1] Arulmozhi D K, Portha B. GLP-1 based therapy for type 2 diabetes[J]. European Journal of Pharmaceutical Sciences, 2006, 28（1-6）：96-108.
[2] 姚艳丽, 冯凭. 胰高血糖素样肽-1 与Ⅰ型糖尿病治疗[J]. 生命的化学, 2005, 25（4）：316-317.
[3] Giorgino F, Laviola L, Leonardini A, et al. GLP-1 ：a new approachfor type 2 diabetes therapy[J]. Diabetes Research and ClinicalPractice, 2006, 74 ：S152-S155.
[4] 李慧, 陈家琪. 胰高血糖素样肽1 类似物的研究进展[J]. 天津药学, 2005, 17（3）：41-43.
[5] Cramer CL, Boothe JG, Oishi KK. Transgenic plants for therapeuticproteins ：Linking upstream and down stream strategies[J]. CurrTopics Microbiol Immunol, 1999, 240 ：95-11.
[6] Staub JU, Garcia B, Graves J, et al. High-yield production of a humantherapeutic protein in tobacco chloroplasts[J]. Nature Biotechnology,2000, 18 ：333-338.
[7] Daniell H, Lee SB, Panchal T, et al. Expression of the native choleratoxin B subunit gene and assembly as functional oligomers in transgenictobacco chloroplasts[J]. J Mol Biol, 2001, 311 ：1001-1009.
[8] Biemelt S, Sonnewald U, Galmbacher P, et al. Production of humanpapilloma virus type 16 virus-like particles in transgenic plants[J].J Virol, 2003, 77 ：9211-9220.
[9] Meyers A, et al. Expression of HIV-1 antigens in plants as potentialsubunit vaccines[J]. BMC Biotech-nology, 2008, 8（53）：1-15.
[10] Abdoreza DS, Nalapalli S, Henry D. The green vaccine ：A globalstrategy to combat in factious and autoimmune diseases[J]. HumVaccin, 2009, 5（7）：488-493.
[11] Horsch RB, Fry JE, et al. A simple and general method of transferringgenes into plants[J]. Science, 1985, 227 ：1229-1231.
[12] Jefferson RA, Kavanagh, Bevan MW. GUS fusion ：beta-glucuronidaseas a sensitive and versatile gene fusion marker in higher plants[J]. EMBO J, 1987, 6（13）：3901-3907.
[13] 王关林, 方宏筠. 植物基因工程原理与技术[M]. 北京：科学出版社,1998.
[14] O’Harte FP, Mooney MH, Lawlor A, et al. N-terminally modifiedglucagon-like peptide-1（7-36）amide exhibits resistance toenzymatic degradation while maintaining its antihyperglycaemicactivity in vivo[J]. Biochimica et Biophysica Acta, 2000, 1474（1）：13-22.
[15] 黄静, 徐进, 左翼, 等. 重组人胰高血糖素样肽-1 的表达及生物学活性[J]. 中国生物工程杂志, 2006, 26（1）：50-55.
[16] Kozak M. Compilation and analysis of sequences upstream from thetranslational start site in eukaryotic mRNAs[J]. Nucleic AcidsRes, 1984, 12（2）：857-872.
[17] 杜小春, 何正权, 陈磊. 植物生物反应器表达药用蛋白研究新进展[J]. 中国生物工程杂志, 2008, 28（9）：135-143.
[18] Yasuda H, Tada Y, Hayashi Y, et al. Expression of the smallpeptide GLP-1 in transgenic plants[J]. Transgenic Research,2005, 14（5）：677-684.
[19] 路宝玉. 烟叶和烟气中化学成分及其危害[J]. 中国公共卫生,1997, 13（6）：322-323.
[20] 杨振泉, 刘巧泉, 焦新安. 利用转基因植物表达药用蛋白[J].中国生物工程杂志, 2004, 24（3）：22-25.