山西农业大学学报（自然科学版）2016，Vol.36Issue(6)：381-386,438,7.
玉米磷酸丙糖异构酶基因的克隆及原核表达
Cloning and prokaryotic expression of TPI gene in maize
摘要
Abstract
ZmT PI gene cloned from Zea mays was expressed in E .coli Bl21. In this work ,based on the full‐length cD‐NA sequences of maize T PI gene ,a cDNA coding region fragment about 750 bp was amplified using RT‐PCR method from total RNA of maize leaves and cloned into the prokaryotic expression vector PGEX ‐4T‐1 ,resulting the recombi‐nant plasmid of GST‐ZmTPI .The resulting construct GST‐ZmTPI was then transformed into E .coli Bl21 for further analysis .Results showed that the ZmT PI (GenBank :EU959275) cDNA sequence is 1 216 bp in length and has an open reading frame (ORF) of 753 nucleotides ,encoding a protein of 250 amino acids with a predicted molecular mass of approximately 26.7205 kDa polypeptide and an isoelectric point of 5.12 ;The ZmTPI protein has highly conserved TIM domain ,and was highly similar with its homologous proteins in other plants ;phylogenetic analysis showed that the de‐duced amino acid sequence of ZmTPI was most similar to that of Saccharum o f f icinarum ,indicating that they belong to the same evolutionary branch ;SDS‐PAGE assay showed that the fusion protein was successfully expressed and puri‐fied with the expected size .The ZmT PI gene was cloned and successfully expressed and purified in E .coli Bl21. This study will provide a foundation for further research on biological function of this protein .关键词
玉米/磷酸丙糖异构酶/基因克隆/原核表达Key words
Maize/Triosephosphate isomerase/Gene cloning/Prokaryotic expression分类
农业科技引用本文复制引用
马芳芳,苏彦冰,张彬,李红英,韩渊怀..玉米磷酸丙糖异构酶基因的克隆及原核表达[J].山西农业大学学报（自然科学版）,2016,36(6):381-386,438,7.基金项目
山西农业大学博士启动基金（2013YJ04）；山西农业大学科技创新基金 （）
