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种子蛋白体靶向表达口服重组胰岛素原载体构建及转化水稻 被引量:1

Construction of Seed Protein Body Targeted Expression Vector and Transformation Rice for Oral Recombinant Proinsulin
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摘要 应用水稻种子生物反应器开发口服重组胰岛素原具有重要应用前景。通过分子设计保证重组胰岛素原在人体肠道内的自主加工成熟,根据水稻密码子偏爱性人工合成了霍乱毒素β亚基和人胰岛素原的融合基因(cholera toxin B subunit fused with human proinsulin,CTBIN),并在C末端添加内质网滞留信号KDEL。通过PCR技术从粳稻品种日本晴全基因组中克隆谷蛋白启动子及其信号肽序列pGluB1sig(GluB1promoter and its signal peptide)用于驱动融合基因CTBIN的表达,插入载体pCAMBIA1302,构建了水稻种子蛋白体靶向表达口服重组胰岛素原的载体pCAMBIA1302-pGluB1sig-CTBINNos。采用农杆菌介导法转化日本晴,获得了46株转基因水稻植株,Western杂交检测到CTB-人胰岛素原融合蛋白在水稻种子中表达。 Developent of oral recombinant proinsulin using rice seeds as bioreactor shows good application future.A fusion gene of the cholera toxin B subunit and human proinsulin(CTBIN)with an endoplasmic reticulum retention signal(KDEL)at C-terminus was designed for its self-maturation process in human gut and synthesized according to the optimized code usage bias of rice.The synthetic CTBIN was introduced to the vector pCAMBIA1302 and expressed under the 2.4kb promoter sequence of rice glutelin GluB1 with its signal peptide(pGluB1sig),which was cloned by PCR from the japonica rice Nipponbare genome.Thus the rice seed protein body targeted expression vector pCAMBIA1302-pGluB1sig-CTBIN-Nos for oral delivery of recombinant proinsulin was constructed and then transformed into Nipponbare via Agrobacterium-mediated method.Totally 46 transgenic rice plants were obtained and the expression of the fusion protein CTB::Human proinsulin in rice seeds was detected by Western-blot.
出处 《中国水稻科学》 CAS CSCD 北大核心 2015年第5期481-489,共9页 Chinese Journal of Rice Science
基金 农业部转基因新品种培育重大专项(2011ZX08010-003) 国家自然科学基金资助项目(30871511)
关键词 口服重组胰岛素原 蛋白体靶向表达 载体构建 转化 水稻 oral recombinant proinsulin protein body targeted expression vector construction transformaiton rice
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  • 1Krauland A H, Guggi D, Bernkop Schnt/rcb A. Oral insulin delivery: The potential of thiolated chitosan insulin tablets on non-diabetic rats. J Contr Rel, 2004, 95(3) : 547 555.
  • 2Wahren J, Ekberg K, J/Srnvall H. C-peptide is a bioactive pep tide. Diabetologia, 2007, 50(3) : 503-509.
  • 3孙学武,袁定阳,谭炎宁,张磊,唐芬,孙志忠,余东,刘瑞芬,段美娟.利用黄瓜做生物反应器表达人胰岛素的研究[J].安徽农业科学,2013,41(12):5201-5203. 被引量:2
  • 4Kwon K C, Verma D, Singh N D, et al. Oral delivery of hu- man biopharmaceutieals, autoantigens and vaccine antigens bioencapsulated in plant cells. Adv Drug Del Rev, 2013, 65 (6) : 782-799.
  • 5Nitesh S C, Sanjeev C, Vandana H, et al. Recent advances in insulin delivery systems: An update. World Appl Sci J, 2010, 11(12): 1552 1556.
  • 6Obembe O O, Popoola J O, I.eelavathi S, et al. Advances in plant molecular farming. Biotechnol Adv, 2011, 29(2): 210 222.
  • 7Xie T, Qiu Q, Zhang W, et al. A biologically active rhIGF 1 fusion accumulated in transgenic rice seeds can reduce blood glucose in diabetic mice via oral delivery. Peptides, 2008, 29 (11): 1862 1870.
  • 8Wakasa Y, Takagi H, Hirose S, et al. Oral immunotherapy with transgenic rice seed containing destructed Japanese cedar pollen allergens, Cry j 1 and Cry j 2, against Japanese cedar pollinosis. Plant Biotechnol J , 2013, 11(1): 66-76.
  • 9Katsube T, Kurisaka N, Ogawa M, et al. Accumulation of soybean glycinin and its assembly with the glutelins in rice. Plant Physiol, 1999, 120(4): 1063-1074.
  • 10Takagi H, Saito S, Yang L, et al. Oral immunotherapy a- gainst a pollen allergy using a seed-based peptide vaccine. Plant Biotechnol J, 2005, 3(5): 521-533.

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