海雀稗酵母单杂文库构建及Pvrboh5上游调控因子筛选

打开文本图片集

中图分类号：S688.4 文献标识码：A 文章编号：1007-0435（2026）02-0447-09

Abstract：To explore the molecular regulatory mechanisms of Purboh5 expression，this study constructeda yeast one-hybrid library from P . vaginatum （Paspalum vaginatum Swartz） using Gateway technology and screened the upstream regulatory factors of Purboh5. Salt-treated stolons of P . vaginatum‘Sea Isle 2000' were used as experimental material.The promoter sequence of Purboh5 was amplified by PCR and subsequently ligated into the yeast one-hybrid bait vector pBait-AbAi，the construct Was named as pPurboh5-AbAi. This construct was then transformed into the Y1H Gold yeast strain to generate the recombinant bait strain. Concurrently，the yeast one-hybrid library was constructed using Gateway technology.Following mRNA isolationand purification，three reading-frame double-stranded cDNAs were synthesized.Primary libraries were constructed via BP reaction，，followed by the generation of high-quality secondary libraries through LR reaction. Finally，transcription factors interacting with Purboh5 were identified by screening the P .vaginatum yeast cDNA library. The results showed that transcriptional autoactivation of the pPurboh5-AbAi/Y1H Gold bait strain was effectively suppressed at an AbA concentration of 100ng⋅mL-1 . In the screening experiment，a total of 18 proteins that bind to the Purboh5 promoter were identified.These findings provide a theoretical foundation for further elucidating the regulatory mechanisms of the Purboh5 gene.

Key words： Paspalum vaginatum; Yeast one-hybrid; Gateway technology; Porboh5 promoter; Transcription factor

海雀稗（PaspalumvaginatumSwartz），又称夏威夷草（Seashorepaspalum），是禾本科（Gramineae），雀稗属（Paspalum）的一种多年生暖季型草坪草，主要分布在热带和亚热带地区，是世界公认的暖季型优质草坪草。（剩余16578字）