生物技术通报2025，Vol.41Issue(12)：66-73,8.DOI:10.13560/j.cnki.biotech.bull.1985.2024-1133
一种可视化筛选转基因通用载体pCamRUBY的构建和应用研究
Construction and Application Study of a General Vector pCamRUBY for Visual Screening of Transgenes
摘要
Abstract
[Objective]To construct and verify an universal vector for visual screening transgenic plants and improving the efficiency of plant genetic transformation.[Method]Having gene RUBY as the reporter gene,a universal expression vector pCamRUBY was constructed using 2A technology.Validation vectors pCamSpeRUBY and pCamKanRUBY were constructed using Spe and Kan resistance genes,respectively.Arabidopsis and cotton were used as the test materials for genetic transformation and pigment content identification.[Result]This vector has the characteristics of high visualization,small vector sequence,MCS sites composed of Sal I,Apa I,Xba I,Sac I and Spe I,and T2A(optimized)sequences.The red positive transgenic Arabidopsis and cotton plats and tissues were visually screened with the naked eye respectively,and stable genetic inheritance was obtained.[Conclusion]The universal expression vector pCamRUBY can directly be linked to the normal exogenous genes,and the positive transgenic plants show red character.The transgenic plants can be directly observed and screened with naked eye,and not affect other phenotypic traits including chlorophyll content of positive plants.关键词
遗传转化/通用载体/报告基因/RUBYKey words
genetic transformation/universal vector/reporter gene/RUBY引用本文复制引用
DOU Shuo,WU jin-hua,WU Li-zhu,DING Ruo-xi,SUN Xing,GUO Wen-jing,KONG Wen-hui,YUAN Jing-xian,ZHANG Dong-mei,WANG Xing-fen,MA Zhi-ying..一种可视化筛选转基因通用载体pCamRUBY的构建和应用研究[J].生物技术通报,2025,41(12):66-73,8.基金项目
国家自然科学基金项目(32072109),河北省中央引导地方科技发展资金项目(236Z2908G) （32072109）
