CAJ | 학술논문

将从胡杨中克隆的果糖-1,6-二磷酸醛缩酶基因(PeALD)构建到pGEX-4T-1载体上，经IPTG诱导成功获得融合蛋白GST：PeALD，并将蛋白进行纯化，其大小约为52kD，转化大肠杆菌的耐盐性实验表明，PeALD基因的成功表达有利于提高大肠杆菌菌株的耐盐性。为研究该基因编码蛋白在植物细胞中的定位，将基因的ORF区构建到定位表达载体pMDC85上，通过PEG介导的拟南芥瞬时转化法观察融合蛋白PeALD：GFP在细胞中的定位情况，结果显示该蛋白定位于胞质中，由此说明实验克隆得到的该胡杨果糖-1,6-二磷酸醛缩酶基因编码的是胞质蛋白。烟草种子在盐培养基上的萌发实验结果显示转基因烟草具有更高的耐盐性；烟草水培苗经200mmol/LNaCl处理一周后，可溶性糖的质谱检测结果显示转基因植株中葡萄糖的含量有很大提高。表明胡杨果糖-1,6-二磷酸醛缩酶通过促进糖酵解和有氧呼吸途径来提高植物对盐胁迫的适应性。
장종호양중극륭적과당-1,6-이린산철축매기인(PeALD)구건도pGEX-4T-1재체상，경IPTG유도성공획득융합단백GST：PeALD，병장단백진행순화，기대소약위52kD，전화대장간균적내염성실험표명，PeALD기인적성공표체유리우제고대장간균균주적내염성。위연구해기인편마단백재식물세포중적정위，장기인적ORF구구건도정위표체재체pMDC85상，통과PEG개도적의남개순시전화법관찰융합단백PeALD：GFP재세포중적정위정황，결과현시해단백정위우포질중，유차설명실험극륭득도적해호양과당-1,6-이린산철축매기인편마적시포질단백。연초충자재염배양기상적맹발실험결과현시전기인연초구유경고적내염성；연초수배묘경200mmol/LNaCl처리일주후，가용성당적질보검측결과현시전기인식주중포도당적함량유흔대제고。표명호양과당-1,6-이린산철축매통과촉진당효해화유양호흡도경래제고식물대염협박적괄응성。
Thefructose-1,6-bisphosphatealdolasegene(PeALD)ofPopulus euphraticawasclonedandconstructed to an expression vector pGEX-4T-1. And then the recombinant plasmid pGEX-PeA LD was transformed into E. coli competent cell BL21 (DE3) to identify the expression of fusion protein GST:PeALD that induced by IPTG . The SDS-PAGE gel electrophoresis showed that the molecular weight of protein coded by PeA LD gene was about 52 kD. The E. coli strains expressing the PeALD showed a higher salt tolerance compared to control strains that expressing the null vector. We constructed the fusion expression vector PeALD:GFP and identify the localization of PeALD in A rabidopsis mesophyll protoplast. The green fluorescence detected by fluorescence microscope showed that the protein encoded by PeA LD localized in the cytoplasm. Seed germination experiment showed that genetically modified tobacco seeds exhibited a higher salt tolerance. The content of soluble sugars in wild-type and transgene lines was analyzed after salt treatment. Results show that the content of glucose in transgenic plants was greatly elevated under salt stress. Our data suggest that PeALD enhanced plant salt adaptation through up-regulation of glycolysis and aerobic respiration.