番茄Slβ-Hex的分子特征及启动子sgRNA分析
Molecular characteristics and promoter analysis of Slβ-Hex gene in tomato
摘要
Abstract
[Objective]Playing an important role in fruit ripening and softening,In order to inactivate the tomato cell wall glycoside hydrolase gene β-Hex by mutation of CRISPR-Cas system and inhibit the degra-dation of n-glycoprotein and the production of free n-glycan,to reduce the degree of hydrolytic softening of cell wall and regulate the excessive softening of tomato fruit lay the foundation for further research.CRISPR-Cas gene editing system will be used to regulate tomato fruit softening by site-specific mutation.[Methods]The characteristics,conserved domain,genome structure,digital expression profile and promoter cis-acting elements of Slβ-Hex polypeptide were systematically analyzed using biological online tools,and suitable sgRNAs were designed and screened according to CRISPR-Cas9 target design principles to promote the con-struction of tomato gene editing efficient expression vector.[Results]Tomato SLβhex gene was located on the complementary chain of tomato chromosome 1,and composed of 2 exons and 1 intron,encoding 575 amino acids.Transcriptome analysis of Slβ-Hex based on RNA-seq showed that Slβ-Hex was mainly expressed in the fruit of tomato,especially in the peel.The positive chain of the promoter distributed 7 Sgrnas,negative chain distributed 20 Sgrnas,of which 10 Sgrnas contained 16 cis-acting elements.[Conclusion]High-specific sgRNA is selected for gene editing to mutate the cis-element sequence and inhibit the expression of Slβ-Hex.关键词
番茄/Slβ-Hex分子特征/启动子/顺式作用元件/sgRNAKey words
tomato/molecular characteristics of Slβ-Hex/promoter/Cis-acting element/sgRNA分类
农业科技引用本文复制引用
张西英,刘江娜,白云凤,李荣霞,张爱萍..番茄Slβ-Hex的分子特征及启动子sgRNA分析[J].新疆农业科学,2025,62(5):1131-1138,8.基金项目
新疆生产建设兵团科技创新人才计划项目"基于双基因编辑调控番茄果实软化研究"(2023CB019-01) （2023CB019-01）
国家自然科学基金项目"基于双基因编辑创制高番茄红素新疆加工番茄新种质"(32160710) （32160710）
新疆生产建设兵团科技创新人才计划项目"加工番茄高可溶性固形物种质创新"(2023CB019-02) （2023CB019-02）
第六师科技计划项目"加工番茄新品种引育及体质增效关键技术研究"(2206) The S & T Innovation Talent Program Project of XPCC"Study on the Regulation of Tomato Fruit Softening Based on Two-Gene Edi-ting"(2023CB019-01) （2206）
National Natural Science Foundation of China"New Germplasm of High Lycopene Xinjiang Processed Tomato Based on Double Gene Editing"(32160710) （32160710）
The S & T Innovation Talent Program Project of XPCC"Processed Tomato Highly Soluble and Solid Material and Germplasm Innovation"(2023CB019-02) （2023CB019-02）
The Sixth Division Science and Technology Program Project"Research on Key Technology of Breeding and Physical Enhancement of New Tomato Varieties"(2206) （2206）
