CAJ | 학술논문

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.
통과체외조작,대강두이단백매억제제(cpti)기인진행수식,획득료일개융합단백기인(sck).해기인시재cpti기인적기출상,재기5'단첨가료신호태편마서렬,재3'단첨가료내질망체류신호편마서렬,지재인도기인전역산물진입세포내질망,병최종체류재내질망급기연생적단백체내.용sck기인전화연초(Nicotiana tabacum L.),대획득적전기인식주진행ELISA검측.결과표명,함유수식기인적전기인연초CpTI단백함량유명현제고,비전미수식cpti기인연초평균고출2배,최고단주가체4배이상,동시전기인식주적항충성야유료현저적제고.결과표명,채용외원단백파향정위적책략,가대폭도제고외원단백재전기인식물세포내적적루량,재식물기인공정연구중구유엄범적차감의의.
Targeting of the synthesized polypeptide in the cells is an important research field in moderncell biology. Cowpea trypsin inhibitor (cpti) gene has been modified and a fusion protein gene (sck) wasproduced by fusing a signal peptide sequence at cpti 5' end and an endoplasm reticulum (ER) retentionsignal peptide at cpti3' end respectively. The signal peptide can direct the newly synthesized polypeptideinto 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 scktransgenic tobacco (Nicotianatabacum L.) was two times higher than cpti transgenic tobaccos and some individuals were four timeshigher. At the same time, sck transgenic tobacco has a high resistance to Lepidoptera pest due to theincreased accumulation level of foreign CpTI protein. The strategy of foreign protein targeting can be usedto increase the accumulation level of foreign protein in transgenic plants and can be widely applied toother related research field in plant genetic engineering.