百合LoTCP4基因克隆分析及过表达载体构建

Cloning and Analysis of LoTCP4 Gene in Lily and Construction of Overex-pression Vector

TCP转录因子在植物整个生命周期过程中发挥着重要的作用,研究TCP基因对于改变植物器官形态辅助分子育种具有重要意义。本研究在百合转录组测序数据库中筛查到一个TCP4基因转录本,利用PCR技术克隆了LoTCP4基因。结果表明,LoTCP4基因包含一个完整的长度为1 038 bp的开放阅读框,编码一个由345个氨基酸组成的疏水性蛋白,包括α-螺旋(18.26%)、延伸链(11.30%)、无规则卷曲(50.25%),β-转角(2.9%)四种二级结构,与拟南芥中AtTCP4基因同源性最高。蛋白质互作预测分析表明,LoTCP4蛋白可能与TCP转录因子家族中其他TCPs蛋白、鳞状细胞启动子结合蛋白(squamosa promoter binding protein-like, SPL)相互作用,GO分析表明LoTCP4主要参与植物花序、芽、茎器官的发育和调节;进一步采用同源重组和golden gate无缝克隆的办法构建了LoTCP4过表达载体。本研究为后续研究LoTCP4基因功能明确其对百合鳞茎发育的影响,提供了一定的科学依据。

TCP transcription factors play an important role in the whole life cycle of plants,and the study of TCP genes is of great significance for changing the morphologyof plant organs to assist molecular breeding.In this study,a TCP4 gene transcript was screened in the lily transcript sequencing database,and the LoTCP4 gene was cloned by PCR technology.The results showed that the LoTCP4 gene contained a complete open readingframe with a length of 1038 bp,encoding a hydrophobic protein composed of 345 amino acids,including four secondary structures:α-he-lix(18.26%),extended strand(11.30%),random coil(50.25%),andβ-turn(2.9%),which had the highest homology with the AtTCP4 gene in Arabidopsis.Predictive analysis of protein interaction showed that LoTCP4 protein mayin-teract with other TCPs proteins and squamosa promoter binding protein-like(SPL)in the TCP transcription fac-tor family,and GO analysis showed that LoTCP4 was mainlyinvolved in the development and regulation ofplant in-florescences,buds and stem organs.Furthermore,the methods of homologous recombination and golden gate seam-less cloning were used to construct LoTCP4 overexpression vectors.This study laid a theoretical foundation for the subsequent studyofthe function of LoTCP4 gene and its effect on the development oflilybulbs.