利用BglBrick方法进行HSA/Exendin-4长效融合蛋白的快速组装

doi:10.13523/j.cb.20140410

中国生物工程杂志

2014, Vol. 34

Issue (4): 59-64 DOI: 10.13523/j.cb.20140410

技术与方法

利用BglBrick方法进行HSA/Exendin-4长效融合蛋白的快速组装

邵妤^1,2, 曲国龙², 金晶^2,3, 王微^2,4, 谭俊杰², 阚乃鹏^1,2, 李玉霞², 刘刚², 陈惠鹏²

1. 安徽大学生命科学学院合肥 230601;
2. 军事医学科学院生物工程研究所北京 100071;
3. 沈阳药科大学沈阳 110016;
4. 吉林大学长春 130012

Rapid Assembly of Long-term HSA/Exendin 4 Fusion Protein via BglBrick Method

SHAO Yu^1,2, QU Guo-long², JIN Jing^2,3, WANG Wei^2,4, TAN Jun-jie², KAN Nai-peng^1,2, LI Yu-xia², LIU Gang², CHEN Hui-peng²

1. College of Life Science, Anhui University, Hefei 230601, China;
2. Institute of Biotechnology AMMS, Beijing 100071, China;
3. Shenyang Pharmaceutical University, Shenyang 110016, China;
4. Jilin University, Changchun 130012, China

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摘要：

目的：通过合成生物学的BglBrick方法快速构建不同种类HSA/ Exendin-4长效融合蛋白，为进行各HSA/Exendin-4融合蛋白生物学活性的筛选比较奠定基础。方法：选用酵母表达载体pPICZαA质粒，构建pPICZαA-E4和pPICZαA-HSA两种基础Brick表达载体，运用BglBrick方法，实现了不同数目Exendin-4分子在HSA的不同末端的快速组装。将构建出的10种HSA/Exendin-4融合蛋白，转入毕赤酵母菌KM71H中，用甲醇诱导目的蛋白的表达。结果：用BglBrick方法构建的HSA/Exendin-4融合蛋白在毕赤酵母中都成功诱导表达出相应的目的蛋白。结论：利用BglBrick方法能够实现HSA长效融合蛋白的快速组装。

关键词： BglBrick; Exendin-4; HSA融合蛋白; 合成生物学

Abstract:

Objective: Different kinds of HSA/Exendin-4 fusion protein are rapidly constructed using BglBrick method of synthetic biology. It lays a foundation for screening and comparison of the bioactivity of fusion protein in each HSA/Exendin-4. Method: Two basic Brick expression vectors, pPICZαA-Exendin-4 and pPICZαA-HSA, are constructed based on the yeast expression vector, pPICZαA plasmid. Rapid assemblies of different amounts of Exendin-4 molecules at the terminals of HSA are achieved via BglBrick method. Ten constructed HSA/Exendin-4 fusion proteins are integrated into the chromosome of Pichia pastoris KM71H, and the corresponding target proteins are expressed after methanol induction. Result:The constructed HSA/Exendin-4 fusion proteins via BglBrick method successfully express the corresponding target protein in Pichia pastoris after methanol induction. Conclusion: The use of BglBrick method can help fulfill the rapid assembly of long-term HAS fusion proteins.

Key words: BglBrick Exendin-4 HAS fusion protein Synthetic biology

收稿日期: 2013-12-24 出版日期: 2014-04-25

ZTFLH:

Q786

基金资助:

“十二五”重大新药创制国家科技重大专项资助项目（2012ZX09301003）

通讯作者: 刘刚 E-mail: jueliu@sohu.com

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引用本文:

邵妤, 曲国龙, 金晶, 王微, 谭俊杰, 阚乃鹏, 李玉霞, 刘刚, 陈惠鹏. 利用BglBrick方法进行HSA/Exendin-4长效融合蛋白的快速组装[J]. 中国生物工程杂志, 2014, 34(4): 59-64.

SHAO Yu, QU Guo-long, JIN Jing, WANG Wei, TAN Jun-jie, KAN Nai-peng, LI Yu-xia, LIU Gang, CHEN Hui-peng. Rapid Assembly of Long-term HSA/Exendin 4 Fusion Protein via BglBrick Method. China Biotechnology, 2014, 34(4): 59-64.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140410 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I4/59

[1] DeFronzo R A, Ratner R E, Han J, et al.Effects of exenatide (exendin- 4) on glycemic control and weightover 30 weeks in metformin - treated patients withtype 2 diabetes. Diabetes Care, 2005, 28(5): 1092.

[2] Kolterman O G, Buse J B, Fineman M S, et al. Synthetic Exendin-4 (AC2993) significantly reduces postprandial and fasting plasma glucose in subjects with type2 diabetes. J Clin Endocrinal Metab, 2003, 88(7): 3082- 3089.

[3] De Leon D D, Crulchlow M F, Ham J Y, et al.Role of glucagon - like peptide - 1 in the pathogenesis and treatment of diabetes mellitus. Int J Biochem Cell Biol, 2006, 38(5~ 6): 845.

[4] 王秀贞,吴军,孟宪军.长效多肽药物合成研究进展.中国生物工程杂志.2003, 23(10):23-27. Wang X Z, Wu J, Meng X J. Progress in the synthesis of long-term peptide medicines. China Biotechnology, 2003, 23(10):23-27.

[5] Chamow S M, Duliege A M, Ammann A, et al. CD4 immunoadhesinsin anti -HIVtherapy:New developments, Int J Cancer, 1992, 7: 69.

[6] Tanaka T,Shiramoto S, Miyashita M, et al. Tumor targeting based on the effect of enhanced permeability and retention (EPR) and the mechanism of receptor mediated endocytosis (RME).International Journal of Pharmaceutics,2004,277:39-61.

[7] Balance D J. 重组人生长激素和人血清白蛋白融合蛋白.英国, CN1207131, 1999. Balance D J. Recombinant Human Growth Hormone and the fusion protein of Human Serum Albumin. Britain, CN1207131, 1999.

[8] Goodey A R. The production of heterologousplasmaproteins. Trends Biotechnol, 1993, 11(10): 430-433.

[9] Yeh P, Landais D, Lemaitre M, et al. Design of yeast-secreted albumin derivatives for human therapy: biological and antiviral properties of a serum albumin-CD4 genetic conjugate. Proc Natl Acad Sci USA, 1992, 89 (5): 1904- 1908.

[10] Osborn B L, Olsen H S, Nardelli B, et al. Pharmacokinetic and pharmacodynamic studies of a human serum albumin-interferon-alpha fusion protein in cynomolgus monkeys. J Pharmacol Exp Ther, 2002, 303 (2): 540- 548.

[11] Sung C, Nardelli B, LaFleur D W, et al. An IFN-beta-albumin fusion protein that displays improved pharmacokinetic and pharmacodynamic properties in nonhuman primates. J Interferon Cytokine Res, 2003, 23 (1): 25- 36.

[12] Canton B, Labno A, Endy D. Refinement and standardization of synthetic biological parts and devices. Nat Biotechnol,2008, 26:787-793.

[13] Endy D. Foundations for engineering biology. Nature, 2005, 438:449-453.

[14] 赵学明,王庆昭.合成生物学:学科基础、研究进展与前景展望.前沿科学,2007,3:56-66. Zhao X M, Wang Q Z. Synthetic biology: Fundamentals advances and prospect. Frontier Science, 2007, 3:56-66.

[15] Hobom B. Gene surgery: on the threshold of synthetic biology. Medizinische Klinik, 1980, 75 (24):834- 841.

[16] 熊燕,陈大明,杨琛等.合成生物学发展现状与前景.生命科学志,2011, 23(9):826-837. Xiong Y, Chen D M, Yang C, et al. Progress and perspective of synthetic biology. Chinese Bulletin of Life Sciences, 2011, 23(9):826-837.

[17] 邢玉华,谭俊杰,李玉霞.合成生物学的关键技术及应用进展.中国医药生物技术.2012, 7(5):357-363. Xing Y H, Tan J J, Li Y X, et al. Key technology and application progress of synthetic biology. Chinese Medicinal Biotechnology, 2012, 7(5):357-363

[18] Anderson J C, Dueber J E, Leguia M, et al. BglBricks: a flexible standard for biological part assembly. J BiolEng, 2010, 4(1): 1-12.

[19] Knight T F. Idempotent vector design for standard assembly of biobricks. DSpace2003http://hdl.handle.net/1721.1/21168.

[20] Shetty R P, Endy D, Knight T F. Engineering BioBrick vectors from BioBrick parts. J Biol Eng,2008, 2(5):1-12.

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