蕙兰CfAP11基因正、反义表达载体的构建及对烟草的转化

Construction of Sense and Antisense Expression Vectors of CfAP11 in C.faberi and Its Transformation to Tobacco

A类基因APETALA1(AP1)属于MADs-box基因,在花的发育中扮演重要的角色,不但可以调控开花时间,而且在ABCDE花发育模型中控制花萼和花瓣的发育。为了研究蕙兰APETALA1基因CfAP11的功能,本研究通过构建蕙兰开花基因CfAP11正义和反义两种植物表达载体,并将其导入农杆菌EHA105菌株,介导遗传转化烟草,对遗传转化的烟草再生植株进行PCR-Southern blot和RT-PCR检测。结果表明,蕙兰CfAP11基因正义和反义表达载体构建成功,并进一步将CfAP11正义基因转化烟草,获得阳性转基因植株。本研究为下一步蕙兰CfAP11基因的功能鉴定提供了实验依据。

APETALA 1 of A class gene belongs to MADS-box gene family,plays an important role in flower development.AP1 gene is the identification gene of flower meristem,and regulates the inflorescence meristem to floral meristem.AP1 gene is the gene of floral organ morphogenesis in the ABCDE model of flower development,and necessary for the normal development of sepals and petals.To characterize the function of CfAP11 gene from Cymbidium faberi,sense and antisense plant expression vectors were constructed and transferred into Agrobacterium tumefaciens strain EHA105.Then vectors were transformed into tobacco plants by using Agrobacterium-mediated method.Regenerated plants of tobacco were detected by PCR-southern blot and RT-PCR.The results showed that the sense and antisense expression vectors were successfully constructed,and the CfAP11 sense gene was further transformed into tobacco.Transgenic plants were obtained successfully.This study provided experimental basis for the next step of functional identification of CfAP11.