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以G418为筛选标记的极细链格孢菌(Alternaria tenuissima)原生质体转化体系的建立(英文) 总被引:2,自引:0,他引:2
[Objective] The study aimed to establish a protoplast transformation system in Alternaria tenuissima. [Method] The protoplast of A.tenuissima was firstly prepared by enzymolysis method; then the yielded protoplast was transformed by G418 resistant DNA plasmid using PEG/CaCl2 method. [Result] The growth phenotype and PCR detection showed that resistance gene had integrated into A.tenuissima genome. The transformation efficiency of this method reached per μg DNA 3-4 transformants. After subculture thrice under nonselective condition, G418 resistance could still inherit stably. [Conclusion] The transformation system of A.tenuissima was successfully established, which laid basis for studying of the gene function of Alternaria tenuissima. 相似文献
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以G418为筛选标记的极细链格孢菌(Alternaria tenuissima)原生质体转化体系的建立 总被引:1,自引:0,他引:1
[目的]建立极细链格孢菌(Alternaria tenuissima)的原生质体遗传转化体系。[方法]采用酶解法制备极细链格孢菌的原生质体,并通过PEG/CaCl2介导的化学方法将含有G418抗性标记的DNA转入极细链格孢菌原生质体。[结果]转化子生长表型及其基因组DNA的PCR检测表明抗性基因已成功整合到极细链格孢菌基因组中。该方法的转化率达3~4个转化子/μg转化DNA。所获得的转化子在无选择压力下连续培养3代,G418抗性仍能稳定遗传。[结论]成功建立了极细链格孢菌的遗传转化系统,为极细链格孢菌的基因功能研究奠定了基础。 相似文献
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