摘要
为进一步提高大豆根瘤菌的固氮能力,以费氏中华根瘤菌(Sinorhizobium fredii)15067基因组的DNA作为模板,采用PCR扩增技术,克隆四碳二羧酸转移酶结构基因dctA的全长序列;将其连入lac启动子下游,构建带有LuxAB基因标记的pTR-Plac-dctA重组表达载体。利用三亲本杂交技术将表达载体转入费氏中华根瘤菌中,构建大豆根瘤菌工程菌株。研究表明,转基因工程菌株侵染大豆幼苗后与出发菌相比,固氮酶活性有了显著提高,为提高大豆产量的研究提供一定参考。
In order to promote the ability of nitrogen fixation, the dctA gene copies in rhizobium fredii were increased. PCR technology was used to clone dicarboxylate transferase gene dctA with template of genomic DNA from Sinorhizobiumfredii 15067. dctA gene was joined into downstream of lac promoter. And expression vector pTR - Plac - dctA was constructed by using plasmid pTR102 marked by luxAB gene. The expression vector pTR -Plac - dctA was transform into Sinorhizobiumfredii 15067 with method of triparental hybridization to construct genetic engi- neering strain. The results showed that the nitrogen fixation ability of genetic engineering strain infect soybean seedlings more increase obviously than original strain. It will lay a foundation for the study of genetic engineering strain influence on nitrogen fixation ability of soybean.
出处
《黑龙江大学自然科学学报》
CAS
北大核心
2012年第3期391-394,400,共5页
Journal of Natural Science of Heilongjiang University
基金
黑龙江省科技攻关资助项目(GA08B101)
哈尔滨市科技创新人才研究专项资金资助项目(RC2009XK001004)
黑龙江大学高层次人才(创新团队)支持计划资助项目(Hdtd2010-05)
