Increasing Accumulation Level of Foreign Protein in Transgenic Plants Through Protein Targeting  

通过细胞内的靶向定位大幅度提高外源蛋白在转基因植株的积累水平(英文)

作　　者：邓朝阳[1] 宋贵生[1] 徐军望[1] 朱祯[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《Acta Botanica Sinica》2003年第9期1084-1089,共6页Acta Botanica Sinica（植物学报：英文版）

基　　金：国家高技术研究发展计划"863"项目(2001AA212041;2001AA222251);国家自然科学基金 (39989001); 国家转基因植物研究与产业化专项 (J99-B-004;J99-A-020);美国洛氏基金(RF97001#581)。~~

摘　　要：Targeting of the synthesized polypeptide in the cells is an important research field in modern cell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) was produced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retention signal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptide into ER, while ER retention signal can make the protein retained in the ER and its derivative protein body. ELISA test indicated that the accumulation level of foreign CpTI protein in sck transgenic tobacco (Nicotiana tabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four times higher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to the increased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be used to increase the accumulation level of foreign protein in transgenic plants and can be widely applied to other related research field in plant genetic engineering.通过体外操作,对豇豆胰蛋白酶抑制剂(cpti)基因进行修饰,获得了一个融合蛋白基因(sck)。该基因是在cpti基因的基础上,在其5'端添加了信号肽编码序列,在3'端添加了内质网滞留信号编码序列,旨在引导基因转译产物进入细胞内质网,并最终滞留在内质网及其衍生的蛋白体内。用sck基因转化烟草(Nicotiana tabacum L.),对获得的转基因植株进行ELISA检测。结果表明,含有修饰基因的转基因烟草CpTI蛋白含量有明显提高,比转未修饰cpti基因烟草平均高出2倍,最高单株可达4倍以上,同时转基因植株的抗虫性也有了显著的提高。结果表明,采用外源蛋白靶向定位的策略,可大幅度提高外源蛋白在转基因植物细胞内的积累量,在植物基因工程研究中具有广泛的借鉴意义。

关 键 词：targeting protein ER localization modified gene Cowpea trypsin inhibitor transgenic tobacco pest resistance analysis 

分 类 号：Q943.2[生物学—植物学] S572[农业科学—烟草工业]