Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (3): 299-307.DOI: 10.11983/CBB19217

• EXPERIMENTAL COMMUNICATIONS • Previous Articles     Next Articles

Cloning and Expression Analysis of Different Truncated U3 Promoters in Phyllostachys edulis

Huijin Fan1,2,Kangming Jin1,Renying Zhuo1,Guirong Qiao1,*()   

  1. 1The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou 311400, China
    2Nanjing Forestry University, Nanjing 210037, China
  • Received:2019-11-07 Accepted:2020-03-24 Online:2020-05-01 Published:2020-07-06
  • Contact: Guirong Qiao

Abstract: The U3 and U6 promoters with well-defined transcription initiation sites are important elements driving sgRNA transcription in the CRISPR/Cas9 genome editing system. According to the two sequences of PeU3 promoter cloned from Phyllostachys edulis, six different truncated U3 promoters were successfully cloned and were 550 bp, 397 bp, 149 bp, and 561 bp, 392 bp, 152 bp, respectively in length. GUS and LUC expression vectors were constructed by corresponding truncated promoter and transformed into the callus of Dendrocalamus latiflorus and tobacco leaf by the Agrobacterium-mediated method, respectively. Our results indicate that, all of these U3 promoters have different transcriptional activity, and the Peu3-1-2pro promoter with a length of 397 bp has the strongest activity. It provides more ideal endogenous promoters for constructing CRISPR/Cas9 genome editing system of P. edulis.

Key words: genome editing, LUC activity, PeU3 promoter, Phyllostachys edulis