作物学报2016，Vol.42Issue(12)：1798-1804,7.DOI:10.3724/SP.J.1006.2016.01798
植酸酶phyA基因的密码子优化及其在大豆中的表达
Codon Optimization and Expression ofphyA Gene in Soybean (Glycine maxMerr.)
摘要
Abstract
Phytic acid is the main anti-nutritional components in the plant origin food. Reducing the phytic acid content of trans-genic plants is an effective way to improve the nutrient utilization rate of soybean. In this research, the codons ofAspergillus ficuumphyA were optimized according to the codon usage bias in soybean. ThephyA(b)gene with scheming DNA sequence was synthesized chemically, which was suitable for expression in soybean. The plant expression vector pCBPS-phyA(b)was con-structed. In the vector,phyA(b) gene was driven by promoter of soybean lectin gene and signal peptide sequence, and transformed into Jilin 35 viaAgrobacterium-mediated method. PCR detection indicated that the target gene was successfully integrated into soybean genome. The protein product of bar gene could be detected in all positive plants by LibertyLink strip analysis. Herbicide leaf painting showed that leaves of wild-type plants were lesioned, while there of transgenic plants remained green. In total, 13 phyA transgenic plants and 19phyA(b) transgenic plants were verified by semi quantitative RT-PCR. The detection results of phy-tase activity, inorganic phosphorus content and phytic acid content in T3 transgenic soybean seeds, showed that the artificial phyA(b) gene was successfully expressed in soybean, and the phytase activity inphyA(b)gene transforming plants was signifi-cantly higher than that inphyAgene transforming plants. It is suggested that codon optimization can significantly improve the expression of foreign genes.关键词
大豆/phyA基因/密码子优化/遗传转化Key words
Soybean/phyA gene/Codon optimization/Genetic transformation引用本文复制引用
寇莹莹,宋英今,杨少辉,王洁华..植酸酶phyA基因的密码子优化及其在大豆中的表达[J].作物学报,2016,42(12):1798-1804,7.基金项目
本研究由国家转基因生物新品种培育重大专项(2014ZX0800404B)资助。This work was supported by the National Transgenic Major Project of China (2014ZX0800404B) （2014ZX0800404B）
