棉花学报2012，Vol.24Issue(4)：285-292,8.
棉花 Gh14-3-3L2基因的分子克隆及其互作蛋白质的初步鉴定
Molecular Cloning of Gh14-3-3L2 and Identification of its Protein Interactions in Upland Cotton
摘要
Abstract
From an EST sequence identified by previous proteomic analysis in our lab we cloned a full-length cDNA, named Ghl4-3-3L2, which encodes a cotton (Gossypium hirsutum) 14-3-3 protein that probably plays a regulatory role in cotton fiber initiation and elongation. To unravel the target gene networks of this protein, the interactome of Ghl4-3-3L2 was studied using the yeast two-hybrid system and a Pull-down assay. However, this protein was severely toxic to yeast. Thus, this paper focuses on the Pull-down results. Six x His tagged Ghl 4-3-3L2 was prokaryotically expressed in E. coli and purified for use as bait protein. Ghl4-3-3L2 associated prey proteins were isolated and enriched from mixed protein samples derived from ovules 3, 1 or 0 days before anthesis and fibers 3 or 6 days after anthesis of wild-type upland cotton cv. Xuzhoul42, the lintless mutant Xuzhoul42, and the Ligon lintless 1 mutant using Pull-down technology. The resulting proteins were further isolated by 2-DE and identified by MALDI-MS/MS analysis. Seven GM4-3-3L2 associated proteins were identified. The putative functions of these proteins included molecular chaperoning, vacuolar trafficking, metabolism, and signal transduction. Some of these proteins, including annexin, vacuolar H+-ATPase and Ran GTPase 3, have been previously linked with fiber development. Our study provides novel data for further elucidation of the molecular functions of Ghl4-3-3L2 and the regulatory mechanisms governing cotton fiber development.关键词
棉花/14-3-3/蛋白质相互作用/Pull-down分析Key words
cotton/ 14-3-3/ protein-protein interaction/ Pull-down assay分类
农业科技引用本文复制引用
惠颖,王晋,孙敬,刘康,唐灿明..棉花 Gh14-3-3L2基因的分子克隆及其互作蛋白质的初步鉴定[J].棉花学报,2012,24(4):285-292,8.基金项目
国家863计划项目(2008AA10Z101) （2008AA10Z101）
转基因生物新品种培育重大专项(2009ZX08009-113B) （2009ZX08009-113B）
