玉米异戊烯基焦磷酸异构酶基因的克隆及其功能分析

Cloning and Functional Analysis of an Isopentenyl Diphosphate Isomerase Gene from Maize

以玉米嫩叶为材料,采用RT-PCR技术克隆了植物萜类物质前体生物合成过程最后一个关键酶——异戊烯基焦磷酸异构酶(IPI)基因的全长cDNA,命名为ZmIPI(GenBank登录号为AY738148),该基因cDNA全长为1 382 bp,包含1 104 bp的开放阅读框,编码367个氨基酸残基.颜色互补分析表明ZmIPI能推动工程菌XL1-Blue+pTrcZmIPI+pAC-BETA超量表达β-胡萝卜素,证实ZmIPI具有典型的IPI基因的功能.

Isopentenyl diphosphate isomerase （IPI） is the last key enzyme of DXP/MVA pathway in biosynthesis of plant terpenoid precursors. The coding sequence of IPI gene was cloned from Zea mays by reversetranscription polymerase chain reaction, which was designated as ZmIPI （GenBank accession No： AY738148）. The full-length cDNA of ZmIPI is 1 382 bp containing a 1 104 bp open reading frame （ORF） encoding a 367-amino-acid. Color complementation assay indicated that ZmIPI could promote the β-carotene accumulation in engineered XL1-Blue haboring pTrcZmIPI and pAC-BETA,and as a result the engineered bacteria showed the brightly orange given by β-carotene. This suggested that ZmIPI had the typical function of known IPI genes.