Patatin Class I与Camv 35s启动子在马铃薯降钙素基因表达中的比较卡达尔和马弗纳)

Fatemeh Ghorbaniparsa, H. Ofoghi
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引用次数: 2

摘要

降钙素(CT)是一种32个氨基酸的多肽激素,是一种强大的特异性骨吸收抑制剂,用于治疗多种人类疾病,如高钙血症和骨质疏松症。迄今为止,许多源自哺乳动物的药用蛋白已经在植物中合成。为了提高植物中异源蛋白的产生水平,选择更强的启动子和优化密码子的使用等策略是人们关注的主要问题。本研究利用农杆菌介导的转化技术,在两种马铃薯品种Kardal和Marfona植株上表达了一个人类降钙素(hCT)基因,该基因由两种不同的启动子(Patatin Class I和花菜花叶病毒35S)驱动。采用分子分析方法对转基因植株进行分析,采用定量EASIA法测定hCT浓度。结果表明,在卡达尔马铃薯块茎中定位产生hCT,总可溶性蛋白为0.7%,而在CaMV 35S启动子中定位产生的总可溶性蛋白为0.2%,在使用Patatin I类启动子和花椰菜花叶病毒35S启动子时定位产生的总可溶性蛋白为0.3%,在总可溶性蛋白中定位产生的总可溶性蛋白为0.1%。数据采用SPSS软件进行分析统计。使用两种不同的启动子和两种不同的品种,hCT生产的平均得分有显著差异。结果表明,在马铃薯中,器官特异性表达导致hCT的积累量比构成型表达量高出近3倍,卡达尔品种的hCT表达量比马弗纳品种高出约2倍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparing Patatin Class I and Camv 35s Promoters in Expression of Human Calcitonin Gene in Potato (Solanum Tuberosum Cvs. Kardal And Marfona)
Calcitonin (CT), a 32 amino acid polypeptide hormone is a powerful and specific inhibitor of bone resorption and is used to treat several human diseases like hypercalcemia and osteoporosis. To date, many pharmaceutical proteins of mammalian origin have been synthesized in plants. To increase the production level of heterologous proteins in plants, strategies such as choice of stronger promoters and optimization of codon usage are of major concern. In this study, a human calcitonin (hCT) gene, driven by two different promoters (Patatin Class I and Cauliflower mosaic virus 35S) was expressed in two types of potato's cultivars (cvs) Kardal and Marfona plants, using Agrobacterium-mediated transformation. The transgenic plants were analyzed by molecular methods and hCT concentration was determined by quantitative EASIA. The results showed the localization of hCT production in Kardal potato tubers led to 0.7% of total soluble proteins whiles total soluble protein was 0.2% when CaMV 35S promoter was deployed, these results of Marfona when used Patatin Class I and Cauliflower mosaic virus 35S promoters were 0.3% of the total soluble protein and 0.1% of the total soluble protein in the respect. Data were analyzed by SPSS software using analytical statistics. There was significant difference in mean score of hCT production when two different promoters and two different cultivars were used. These results showed that organ specific expression in potato led to nearly 3 fold higher hCT accumulation than constitutive expression and Kardal cultivar expressed hCT about 2 times higher than Marfona cultivar.
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