植酸酶基因在具有种子特异性启动子27kDaγZein和组成型启动子CaMV35S的转基因玉米中的表达

Ririn Septina Arthasari, R. A. Wulandari, P. Basunanda
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引用次数: 1

摘要

植酸酶被应用于动物饲料中,以帮助动物吸收更多的营养。使用含有植酸酶的饲料原料有望降低动物饲料生产成本。通过改进遗传学,以组装含有高植酸酶含量的转基因植物的方式,努力提高玉米中植酸酶含量。27kDaγZein启动子是在颖果中表达基因的特异性启动子,启动子CaMV 35S是控制所有组织中基因表达的组成型启动子,通常不依赖于生长阶段。采用根癌农杆菌感染法将转基因玉米转化到玉米B104上。采用逆转录聚合酶链式反应(RT-PCR)方法检测植酸酶基因在授粉后10、20和30天叶片、根和颖果中的表达。植酸酶活性测试也使用比色磷钼酸分析法进行,以单位µg-1的植酸酶活性。结果表明,具有27kDaγZein启动子的转基因植株中的植酸酶基因在玉米颖果中高表达,但在Z6.10系中也在叶片中表达,而在CaMV 35S启动子中,植酸酶基因仅在叶片上表达。植酸酶活性表明转基因玉米高于非转基因玉米。
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Expression of Phytase gene in transgenic maize with seed-specific promoter 27-kDa γ Zein and constitutive promoter CaMV 35S
Phytase enzymes are applied to animal feed to help animals absorb more nutrients. The use of feed raw materials containing phytase enzymes is expected to reduce the cost of animal feed production. Efforts to increase the phytase content in maize were carried out by improving genetics, in the way of assembling transgenic plants containing high phytase content. The 27-kDa γ Zein promoter is a specific promoter that expresses genes in caryopsis, and promoter CaMV 35S is a constitutive promoter that controls gene expression in all tissues and generally does not depend on the growth phase. Transgenic maize was transformed using Agrobacterium tumefacien infection method on maize B104. The reverse transcriptase polymerase chain reaction (RT-PCR) approach was used to examine the expression of phytase genes in leaves, roots, and caryopsis was done 10, 20, and 30 days after pollination (DAP). The phytase enzyme activity test was also carried out by using the colorimetric phosphomolybdate analysis method to see the phytase enzyme activity in unit µg-1. The results showed that the phytase gene in transgenic plants with the 27-kDa γ Zein promoter was highly expressed in maize caryopsis, but in line Z6.10 was also expressed in leaves, while in the CaMV 35S promoter the phytase gene was only expressed on the leaves. Phytase enzyme activity showed that transgenic maize was higher than non-transgenic maize.
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