转PRSV复制酶基因番木瓜食品安全性的初步评价
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阮小蕾,女,1973年生,博士,讲师. 研究方向:植物病理学. E-mail:ruanxl@scau.edu.cn

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国家自然科学基金项目(30370929)资助


Preliminary Evaluation of Food Safety of Transgenic Papaya with the Replicase Gene of PRSV
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    摘要:

    应用生物信息学的方法对转基因番木瓜进行实质等同性分析。将复制酶基因与多个数据库中的花生、大豆、坚果、牛奶、鸡蛋、鱼类、贝类和小麦等8类过敏源序列进行比对分析,结果未发现连续8个相同的氨基酸, 据此可初步认为转入的复制酶蛋白不是已知的致敏源。将复制酶基因进行原核表达,并纯化复性表达蛋白,再通过模拟胃肠液消化试验,结果发现纯化的变性蛋白与复性蛋白均能在15 s内降解,表明复制酶蛋白能在人的消化系统内降解,使人体发生过敏的可能性不大。通过液相萃取法,提取转基因与非转基因番木瓜中的一种内源毒物苄基异硫氰酯(benzyl isothiocyanate,BITC),将浓缩后的样品进行气相色谱分析,结果表明BITC的含量在转基因与非转基因番木瓜中差异不显著。

    Abstract:

    Some indexes of food safety of transgenic papaya with the replicase gene of papaya ringspot virus(PRSV) were evaluated in this thesis.By means of biological informatics,the analysis between the sequence transformed to papaya and the allergen sequences from GenBank,EMBL databases,PIR databases,etc. indicated there were not continuously eight same amino acids compared to the eight classes of allergens,including peanut,soybean,nut,milk,egg,fish,shellfish and wheat.It could be inferred that the rep sequence transformed to papaya did not contain the known allergens. The fulllength rep gene of papaya ringspot virus was cloned into expression vector pET-28b(+) and the specific expression product was purified by Ni-NTA resin. Some protein were attempted to be renatured.Through imitated gastric juice and intestinal juice experiment, it had been further proved the protein product of Rep gene could be degraded in human beings’ digestive system,it was unlikely to cause to the human body to be allergic. From the transgenic and no-transgenic papaya,an endogenous poisonous substance called benzyl isothiocyanate(BITC)was respectively extracted by liquid phase extraction,concentrated and sent to China National Analytical Center,Guangzhou. The results of gas chromatography and biological statistical analysis showed that there was no distinct difference in transgenic and no-transgenic papaya.

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阮小蕾,侯燕,李华平.转PRSV复制酶基因番木瓜食品安全性的初步评价[J].华中农业大学学报,2010,29(3):381-386

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  • 收稿日期:2009-10-09
  • 最后修改日期:2009-11-17
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