Identification of transcription factors of nitrate reductase gene promoters and NRE2 cis-element through yeast one-hybrid screening in Nicotiana tabacum

Abstract

Background: This study aimed to identify the transcription factors of nitrate reductase genes (NIA1 and NIA2) promoters and hypothetical cis-element of NRE2. Based on the constructed cDNA library of Nicotiana tabacum K326, a yeast one-hybrid system was established using the Matchmaker® Gold Yeast One-Hybrid Library Screening System from Clontech. The transcription factors of NIA1 andNIA2 promoters and NRE2 cis-elements were screened. Results: After sequencing and bioinformatics analysis, 15 cDNA sequences were identified: 9 for NIA1 (including XP-016503563.1 and NP-001312236.1), 3 for NIA2 (including XP-016510250.1), and 3 for NRE2 (including XM-016576899.1). XP-016503563.1 was annotated in PREDICTED: CRM-domain containing factor CFM3, and NP-001312236.1chloroplastic/mitochondrial-like in Nicotiana tabacum. NP-001312236.1 was annotated in Sulfite oxidase-like of Nicotiana tabacum. XP-016510250.1 was annotated as PREDICTED: uncharacterized protein LOC107827596 in Nicotiana tabacum. XM-016576899.1 was annotated in PREDICTED: Nicotiana tabacum RING-H2 finger protein ATL16-like (LOC107759033). Conclusion: A yeast one-hybrid library was successfully constructed. The identified transcription factors may provide a theoretical basis for the study of plant nitrate reductase.