作物学报2016，Vol.42Issue(9)：1332-1341,10.DOI:10.3724/SP.J.1006.2016.01332
甘蔗捕光叶绿素a/b结合蛋白基因ScLhca3的克隆及表达
Cloning and Characterization of Light Harvesting Chlorophylla/b-Binding Protein Coding Gene (ScLhca3) in Sugarcane
摘要
Abstract
TheLhcagene family in green plants encodes several light-harvesting chlorophylla/b-binding proteins that harvest and transfer light energy to the reaction center of photosystem I (PSI) in photosynthesis. The cDNA sequence ofLhca3gene was firstly obtained from sugarcane leaf full-length cDNA library through sequencing and validated by homology comparison. It was designatedScLhca3 and submitted to the GenBank (accession number: KU215669).ScLhca3contains an 804 bp open reading frame (ORF) and encodes a deduced protein of 267 amino acids, with a molecular weight and pI of 28.91 kD and 8.96, respectively. Bioinformatics analysis showed that ScLhca3 is a hydrophilic non-secretory protein with three transmembrane domains and a chlorophylla/b binding domain. Sequence multi-alignment and phylogenetic analysis demonstrated that the ScLhca3 protein sequence shared a high identity with Lhca3 from other plants, with the specificity of species. The protokaryotic expression vector pGEX-6P-1-ScLhca3 was constructed and expressed inE. colicells under the induction of IPTG. The subcellular localization experiment showed that the ScLhca3 fused with GFP, a reporter protein, was located on chloroplast. Real-time quantitative PCR analysis showed the expression ofScLhca3 had a clear tissue specificity, and was upregulated by CdCl2, ABA, and H2O2, but downregulated by darkness, NaCl, and PEG.关键词
甘蔗/光系统I/捕光叶绿素a/b结合蛋白/实时定量PCRKey words
Sugarcane/Photosystem I/Light harvesting chlorophylla/b-binding protein/Real-time quantitative PCR引用本文复制引用
翟玉山,邓宇晴,董萌,徐倩,程光远,彭磊,林彦铨,徐景升..甘蔗捕光叶绿素a/b结合蛋白基因ScLhca3的克隆及表达[J].作物学报,2016,42(9):1332-1341,10.基金项目
本研究由国家高技术研究发展计划(863计划)项目(2013AA102604),国家自然科学基金项目(31171605,31371688)和国家现代农业产业技术体系建设专项(CARS-20-1-1)资助。This study was supported by the National High-tech Research and Development Program of China (2013AA102604), the National Natural Science Foundation of China (31171605,31371688), and the China Agriculture Research System (CARS-20-1-1) （863计划）
