大豆高赖氨酸基因PM8在烟草中的超量表达及其抗非生物胁迫功能研究

Over-expression of high lysine gene PM8 (soybean) in tobacco and function study of abiotic stress tolerance

应用生物信息学方法在GenBank数据库查找到1条编码大豆LEA3蛋白的高赖氨酸蛋白基因PM8(登录号:Z22872),并利用RT-PCR方法从大豆干种子中克隆得到该基因,序列分析表明,PM8基因与GenBank上的序列存在5个碱基的差异,使编码蛋白的赖氨酸重量比由原来的19.65%增加为19.93%。构建了PM8基因的植物表达载体pEMT-PM8,用农杆菌介导法转化烟草。从转PM8基因的PCR阳性植株中选取5个既抗盐又抗旱的株系进行Real-timePCR检测。结果表明,PM8基因全部超量表达。研究为培育高营养品质且抗逆的转基因作物研究提供了一条新的途径,具有重要的研究价值。

After searching in GenBank database of NCBI, a gene PM8 （Accession number： Z22872） for high lysine content from soybean（Glycine max） was found, and belongs to the family of late embr-yogenesis abundant （LEA3） proteins. In this experiment, PM8 gene was cloned from dry soybean seeds by RT-PCR. Amino acid sequence analysis demonstrated that it existed five bases difference between PM8 and the sequence in GenBank, which raised the lysine content from 19.65% to 19.93%. Plant expression vector pEMT-PM8 was constructed, and then transformed into tobacco by Agrobacterium-mediated method. The result showed that real-time PCR made the PM8 gene overexpressed in all five PCR positive lines with salt and drought tolerance. This study attempted to provide a new approach to cultivate the crops with discovery and use of high nutrition value and osmotic stress tolerance, which had the important research value.