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Bulletin of Botanical Research ›› 2020, Vol. 40 ›› Issue (2): 293-300.doi: 10.7525/j.issn.1673-5102.2020.02.017

• Research Report • Previous Articles     Next Articles

Gene Cloning and Expression Analysis of PDIL1-2 of Ornamental Kale(Brassica oleracea var. acephala)

LI Yang, SHI Ya-Kun, GAO Shi-Ke, Li Xiao-Yu, LAN Xing-Guo   

  1. College of Life Science, Northeast Forestry University, Harbin 150040
  • Received:2019-07-31 Online:2020-03-05 Published:2020-03-06
  • Supported by:
    Heilongjiang Student's Platform for Innovation and Entrepreneurs(201810225338);Fundamental Research Funds for the Central Universities(2572018BD01);National Natural Science Foundation Project(31870300)

Abstract: The BoPDIL1-2 gene was isolated from the stigma of ornamental kale(Brassica oleracea var. acephala) S13-bS13-b homozygotes by RT-PCR and RACE. The BoPDIL1-2 coding region sequence was inserted into the prokaryotic expression vector pET-14b and transformed into the E.coli cell for purification.We prepared the polyclonal antibodies against recombinant BoPDIL1-2 through immune mouse. The expression level of BoPDIL1-2 in the various tissue anddifferent developmental stigmas were detected by Western blot. The deduced amino acid sequence of BoPDIL1-2 shared 97.3% and 85.5% identity with Brassica napus BnPDIL1-2 and Arabidopsis thaliana AtPDIL1-2, respectively. SDS-PAGE results showed that the recombinant BoPDIL1-2 fusion protein about 58 kDa was induced by IPTG.Western blot results revealed BoPDIL1-2 wasspecifically expressed inthe stigma,the expression of BoPDIL1-2 was lower in the early stage of stigma, while it was higher in the stigma at theflowering stage.

Key words: Brassica oleracea var. acephala, PDIL1-2, prokaryotic expression, polyclonal antibody, expression analysis

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