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筛选标记基因是一种用于植物遗传转化后阳性转化子筛选的有效手段,但目前介导标记基因表达均采用组成性启动子.这些启动子虽然在植物遗传转化得到了广泛应用,但由于是组成性表达,在转基因的生物安全性存在标记基因的安全性担忧.为了克服这一缺点,本文从水稻基因组内克隆了水稻半胱氨酸蛋白酶基因(Cyctein protease,CP)的启动子,通过gus转基因的验证,该启动子只在愈伤组织表达,不在水稻根、茎、叶等组织表达,为愈伤组织特异性表达启动子(Callus-specific Promoter,CSP).将该启动子用于介导筛选性标记基因的表达,通过对筛选性标记基因潮霉素磷酸转移酶基因(hpt)的密码子优化,明显提高了CP启动子在水稻转基因选择的转化效率,质粒转化效率和阳性植株率均达到了pCAMBIA1301的35S启动子介导标记基因hpt的选择效果.
Screening marker gene is an effective method for screening positive transformants after plant genetic transformation, but at present, the constitutive promoter is used to mediate the expression of marker genes. Although these promoters have been widely used in plant genetic transformation, Constitutive expression is concerned with the safety of marker genes in the biosafety of transgene.In order to overcome this shortcoming, we cloned the promoter of rice cystatin (CP) from the genome of rice, The promoter was expressed only in callus but not in roots, stems and leaves of rice, and was Callus-specific Promoter (CSP) Through the codon optimization of the selectable marker gene hpt, the transformation efficiency, the transformation efficiency and the positive plant rate of the CP promoter in rice transgenic plants were significantly increased All achieved the 35S promoter-mediated marker gene hpt selection effect of pCAMBIA1301.