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首页> 外文期刊>British Biotechnology Journal >Comparing the Effect of Two Promoters on CassavaSomatic Embryo at Transient GUS Assay Level
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Comparing the Effect of Two Promoters on CassavaSomatic Embryo at Transient GUS Assay Level

机译：在瞬时GUS分析水平上比较两种启动子对木薯体细胞胚的影响

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35S promoter from the Cauliflower Mosaic Virus (pCaMV) is a constitutive promoter commonly used in plant genetic transformation while Cassava Mosaic Virus (pCsVMV) is another promoter which is underutilized. The combination of the two promoters was used to form (pOYE153). The method adopted includes the insertion of a β–glucuronidase reporter gene (UidA) into a promoter cassette comprising the CsVMV promoter. The second construct (pCAMBIA2310) had (pCaMV) used for the selectable marker and gene of interest. This construct was mobilized into Agrobacterium tumefaciens strain LBA4404 and then tested for expression of the UidA gene in transient assays in cassava somatic embryos. After co-cultivation of these Agrobacterium with the plant tissues, histochemical β–glucuronidase (GUS) assays were performed to determine the level of UidA gene expression in transient assays. The results showed that the pCsVMV was able to drive high gene expression of β–glucuronidase reporter gene (UidA) in the transient assays in cassava somatic embryo. Expression of the gene also increases with the increase in the day of co-cultivation and likewise expression of the gene was higher for the sample in the light than the dark.
机译：花椰菜花叶病毒（pCaMV）的35S启动子是植物遗传转化中常用的组成型启动子，而木薯花叶病毒（pCsVMV）是另一个未得到充分利用的启动子。两种启动子的组合用于形成（pOYE153）。所采用的方法包括将β-葡萄糖醛酸苷酶报道基因（UidA）插入包含CsVMV启动子的启动子盒中。第二种构建体（pCAMBIA2310）具有（pCaMV）用于选择标记和目的基因。将该构建体动员到根癌农杆菌菌株LBA4404中，然后在木薯体细胞胚的瞬时测定中测试UidA基因的表达。将这些农杆菌与植物组织共培养后，进行组织化学β-葡萄糖醛酸酶（GUS）测定，以确定瞬时测定中UidA基因表达的水平。结果表明，在木薯体细胞胚的瞬时测定中，pCsVMV能够驱动β-葡萄糖醛酸糖苷酶报道基因（UidA）的高基因表达。随着共培养天数的增加，基因的表达也增加，同样，在黑暗中样品的基因表达也更高。

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