植酸酶phyA基因的密码子优化及其在大豆中的表达

发布时间：2018-04-16 19:07

本文选题：大豆 + phyA基因　；参考：《作物学报》2016年12期

【摘要】：植酸是植物源食品中的主要抗营养成分,降低植酸含量可有效提高大豆的营养利用率。本文根据大豆密码子使用偏好性,对无花果曲霉植酸酶phyA基因进行密码子优化,人工合成了适合在大豆中表达的phyA(b)基因。以p CAMBIA3301为骨架,构建由大豆凝集素基因启动子和信号肽序列调控的植物表达载体pCBPS-phyA(b)。用农杆菌介导法遗传转化吉林35大豆子叶节。PCR检测表明目的基因已初步整合至大豆基因组中;bar试纸条表明所有阳性植株中均能检测到bar基因的蛋白产物;除草剂叶片涂抹显示野生型的叶片出现黄化或枯萎现象,而转基因植株叶片表现正常,具除草剂抗性;以半定量RT-PCR共筛选到13株转phyA和19株转phyA(b)阳性转基因大豆植株。通过对转基因大豆T_3种子中植酸酶活性、无机磷和植酸磷含量等检测,证明人工基因phyA(b)比phyA在大豆种子中所表达的植酸酶具有更高的活性,说明密码子优化有利于提高外源基因的表达。
[Abstract]:Phytic acid in plant derived foods is the main anti nutrients, reduce the content of phytic acid can effectively improve the utilization rate of soybean nutrition. According to the soybean codon preference, codon optimization of Aspergillus ficuum phytase phyA gene was synthesized, suitable for up to phyA in Soybean (b) gene in P CAMBIA3301. Skeleton, expression vector pCBPS-phyA was constructed by soybean lectin gene promoter and signal peptide sequence control plants (b). By the method of genetic transformation of Soybean Cotyledonary Node of Jilin 35.PCR detection indicated that the target gene has been integrated into the soybean genome by Agrobacterium tumefaciens; bar assay showed that all positive plants were protein product of bar gene the detected; herbicide leaf painting shows the wild type leaves appeared chlorosis or wilting, and the leaves of transgenic plants showed normal with herbicide resistance were screened by semi quantitative RT-PCR; 13 strains of phyA and 19 strains of phyA (b) positive transgenic soybean plants. The activity of phytase transgenic soybean seed T_3, detection of inorganic phosphorus and phytate phosphorus content, it is shown that the artificial phyA gene (b) has higher activity than the phyA expression in the soybean seeds phytase that codon optimization to improve the expression of exogenous gene.

【作者单位】：天津大学环境科学与工程学院;
【基金】：国家转基因生物新品种培育重大专项(2014ZX0800404B)资助~~
【分类号】：S565.1;Q943.2

【相似文献】