Journal of Shanghai University(Natural Science Edition) ›› 2019, Vol. 25 ›› Issue (4): 604-611.doi: 10.12066/j.issn.1007-2861.1949

• Research Articles • Previous Articles     Next Articles

Cloning and expression vector construction of E3 ubiquitin ligase PpARI1 gene from juicy peaches

Jie YUAN1, Fangwei MA1, Mengyun LI1, Qing CAO1, Weiwei ZHENG2, Sibao WAN1()   

  1. 1. Shanghai Key Laboratory of Bio-energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China
    2. The Key Laboratory for Quality Improvement of Agricultural Products of Zhejiang Province, College of Agriculture and Food Science, Zhejiang A & F University, Hangzhou 311300, China
  • Received:2017-05-31 Online:2019-08-30 Published:2019-09-04
  • Contact: Sibao WAN E-mail:wansibao@126.com

Abstract:

Fresh juicy peaches were used as materials, and a novel sequence of E3 ubiquitin ligase PpARI1 was obtained using a polymerase chain reaction (PCR) method with the primers designed according to the predicted gene sequence of ARI1 in NCBI. The open reading frame (ORF) box of PpARI1 gene was forward recombined into pCAMBIAy1300, an efficient expression vector, between Xba I site and BamH I site. The sequencing results showed that the obtained PpARI1 was 1 764 bp with 99.83${\%}$ identity to that of the predicted gene sequence, and the ORF of PpARI1 gene was correctly inserted between the promoter and the terminator. The recombined plasmid was transformed to competent Agrobacterium cells with a heat shock method. The PCR result showed that the vector pCAMBIAy1300-PpARI1 was successfully transferred into Agrobacterium. The results would lay a foundation for further function researches of PpARI1 gene in juicy peaches.

Key words: juicy peach, E3 ubiquitin ligase, cloning, expression vector construction

CLC Number: