摘要
以快生型大豆根瘤菌(Sinorhizobium fredii)15067的基因组DNA作为模板,采用PCR技术,克隆结瘤基因nodD编码区序列;利用DNA重组技术,将nodD基因连到lac启动子的下游,通过luxAB基因标记的质粒载体pTR102构建表达载体pTR102-Plac-nodD,采用三亲本杂交的方式,将构建的表达载体转化土著大豆根瘤菌,构建转基因工程菌株,为研究转基因工程菌株对大豆结瘤能力的影响提供依据。
PCR technology was used to clone nodulation gene nodD with template of genomic DNA from Sinorhizobium fredii 15067 in this study. Utilization of DNA recombination technique, nodD gene was joined into downstream of lac promoter. And expression vector pTRlO2-Plac-nodD was constructed by using plasmid pTR102 marked by luxAB gene. The expression vector pTRlO2-Plac-nodD was transformeded into native Rhizobiurn japonicum with method of triparental hybridization to construct genetic engineering strain. It will lay a foundation for the study of genetic engineering strain influence on nodulation ability of glycine max.
出处
《黑龙江大学自然科学学报》
CAS
北大核心
2012年第1期121-123,129,共4页
Journal of Natural Science of Heilongjiang University
基金
哈尔滨市科技创新人才研究专项资金资助项目(RC2009XK001004)
黑龙江省科技攻关项目(GA08B101)
黑龙江大学高层次人才(创新团队)支持计划资助项目(Hdtd2010-05)
关键词
大豆根瘤菌
nodD基因
工程菌株
三亲本杂交
Rhizobium japonicum
nodD gene
genetic engineering strain
triparental hybridization
