CAJ | 학술논문

黄曲霉毒素污染对花生产业危害巨大,通过基因工程改变花生果种皮结构以提高花生抗黄曲霉能力。根据 GenBank 中拟南芥 AtTTG1基因的 cDNA 编码区设计引物,通过 RT-PCR 克隆AtTTG1基因,结果显示,克隆获得的片段长1026bp,与基因库中数据比对相同,该片段编码341个氨基酸,预测其蛋白分子量为86.96KDa,等电点为4.85。结合实验室已获得的花生果种皮特异启动子 S19和 MAR 序列调控的植物表达载体 pLMAR,构建了植物高效表达载体 pLMAR-S19-TTG1,并将其导入根癌农杆菌 EHA105。为进一步对花生进行遗传转化,获得转基因高抗黄曲霉花生奠定基础。
황곡매독소오염대화생산업위해거대,통과기인공정개변화생과충피결구이제고화생항황곡매능력。근거 GenBank 중의남개 AtTTG1기인적 cDNA 편마구설계인물,통과 RT-PCR 극륭AtTTG1기인,결과현시,극륭획득적편단장1026bp,여기인고중수거비대상동,해편단편마341개안기산,예측기단백분자량위86.96KDa,등전점위4.85。결합실험실이획득적화생과충피특이계동자 S19화 MAR 서렬조공적식물표체재체 pLMAR,구건료식물고효표체재체 pLMAR-S19-TTG1,병장기도입근암농간균 EHA105。위진일보대화생진행유전전화,획득전기인고항황곡매화생전정기출。
Aflatoxin contamination has severely threatened the industry of peanut.We want to ap-ply gene engineering to change the structure of peanut's pod and testa to improve the ability of anti-A. flavus .According to the coding region cDNA of the Arabidopsis thaliana AtTTG 1 gene in the Gen-Bank, we designed the primers, and the AtTTG1 gene was cloned by reverse transcription PCR (RT-PCR).The result showed that the length of the cloned segment was 1026bp,it was same as the data of GenBank by compare.This segment coded 341 amino acids, the predicted protein was about 86.96 KDa, and its isoelectric point was about 4.85.The plant high efficient expression vector of pLMAR-S1 9-TTG1 was successfully constructed.It used pod and testa specific promoter S1 9 of peanut and the plant expression vector of pLMAR which was regulated by MAR sequence, and it was transformed in-to agrobacterium EHA105.This work aimed at laying a foundation for future transformation of pea-nut in order to obtaining transgenic peanut with high resistance to A.flavus .