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金属硫蛋白(MT)是一类广泛存在于生物中的低分子量、富含半胱氨酸且可结合重金属的蛋白质,在土壤重金属污染的治理中发挥重要作用。将从柽柳和星星草cDNA文库中克隆的MT基因构建在同一植物表达载体pRCXM上,通过农杆菌介导的叶盘法转化烟草‘龙江911’,获得了抗卡那霉素的转基因植株。PCR-Southern和Northern blot检测证明双价MT基因已整合进烟草基因组中并可正常表达。抗性试验证明双价MT基因的转入提高了转基因烟草对Cd2+和Cu2+的抗性,转基因烟草分别能在300μmol·L-1Cd2+或150μmol·L-1Cu2+的胁迫下正常生长。本研究结果对通过基因工程技术培育植物修复新品种提供了理论基础。
Metallothionein (MT) is a kind of low-molecular-weight, cysteine-rich and heavy metal-binding protein widely found in living organisms and plays an important role in the management of soil heavy metal pollution. The MT gene cloned from Tamarix ramosissima and Phragmites australis cDNA library was constructed on the same plant expression vector pRCXM. The transgenic tobacco plants resistant to kanamycin were obtained by Agrobacterium tumefaciens-mediated leaf disc transformation of ’Longjiang 911’. PCR-Southern and Northern blot showed that the bivalent MT gene has been integrated into the tobacco genome and expressed normally. Resistance experiments showed that the introduction of bivalent MT gene increased the resistance of transgenic tobacco to Cd2 + and Cu2 +, and the transgenic tobacco plants could grow normally under the stress of 300μmol·L-1Cd2 + or 150μmol·L-1Cu2 +, respectively. The results of this study provide a theoretical basis for cultivating new plant species through genetic engineering.