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[目的]研究公农2号紫花苜蓿抗寒基因CAS19在烟草中的表达。[方法]根据苜蓿抗寒基因(CAS19)核酸序列设计1对引物,用RT-PCR扩增出分离CAS19的蛋白基因,并克隆到pMD18-T Vector载体中,再亚克隆到表达载体PBI121,成功构建重组表达质粒PBCAS。经农杆菌介导转基因入烟草,并通过Sounthern-blotting杂交分析检测转基因苗。[结果]CAS19抗寒基因可以在烟草中得以高效表达。[结论]该研究为进一步探明CAS19抗寒基因在烟草中的表达机制提供了理论依据。
[Objective] The research aimed to study the expression of CAS19 gene in alfalfa (Medicago sativa. [Method] A pair of primers was designed based on the nucleotide sequence of alfalfa cold-resistant gene (CAS19). The protein gene isolated from CAS19 was amplified by RT-PCR and cloned into vector pMD18-T and subcloned into expression vector PBI121 Construction of recombinant expression plasmid PBCAS. Agrobacterium-mediated transgene into tobacco and transgenic seedlings were tested by Sounthern-blotting hybridization assay. [Result] The cold-resistant gene CAS19 was highly expressed in tobacco. [Conclusion] This study provided the theoretical basis for further exploration of the expression mechanism of CAS19 cold resistance gene in tobacco.