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为了选取合适的内参基因来分析培养基中前体物诱导及对照的茶愈伤组织中茶氨酸代谢相关基因的差异表达,利用Gen Bank上登录的茶树基因序列以及通过构建文库测序所得的基因(具有完整的阅读框)共7个持家基因设计引物。在分析这些引物的扩增效率和特异性后,测定了它们在茶愈伤组织生长过程中(对照和愈伤培养基中添加含氮外源物的情况下)的表达水平。利用ge Norm和Norm Finder软件分析了这些持家基因的稳定性,确定在该条件下合适的内参基因为β-actin和GAPDH。
In order to select suitable internal reference genes to analyze the differential expression of theanine metabolism related genes in tea callus induced by the precursors in the medium and the control, the gene sequences of the tea tree genes registered in Gen Bank and the genes sequenced by constructing the library (With a complete reading frame) a total of seven housekeeping gene design primers. After analyzing the amplification efficiency and specificity of these primers, their expression levels during tea callus growth (in the case of control and in the addition of nitrogenous excipients in the callus) were measured. The stability of these housekeeping genes was analyzed using ge Norm and Norm Finder software to determine the suitable internal reference genes for this condition as beta-actin and GAPDH.