Acta Entomologica Sinica ›› 2017, Vol. 60 ›› Issue (1): 18-23.doi: 10.16380/j.kcxb.2017.01.003

• RESEARCH PAPERS • Previous Articles     Next Articles

Eukaryotic expression and function analysis of Bombyx mori inhibitor of apoptosis protein (BmIAP) gene

YI Hua-Shan1,2, MA Xian-Ping1, PAN Min-Hui2,*, LU Cheng2,*   

  1. (1. Department of Animal Medicine, Rongchang Campus of Southwest University, Rongchang, Chongqing 402460, China; 2. State Key Laboratory of Silkworm Genome Biology, Chongqing 400715, China)
  • Online:2017-01-20 Published:2017-01-20

Abstract: 【Aim】 Bombyx mori inhibitor of apoptosis protein (BmIAP) has been found in the silkworm, B.mori, and characterized as a protein of the IAP (inhibitor of apoptosis protein) family. This study aims to verify its function as a suppressor of apoptosis in B.mori cells, so as to further study the roles of BmIAP in cell apoptosis. 【Methods】 The transient expression vector named PIZ/V5-BmIAP-dsRed was transfected in the BmN-SWU1 cells of B.mori by liposome, and the mRNA and protein expression levels of BmIAP gene were evaluated by real-time quantitative PCR and Western blot analysis, respectively. The BmN-SWU1 cells were induced by 150 ng/mL actinomycin D for 12, 18 and 24 h, and the relationship between BmIAP protein and cell apoptosis was assayed by immunofluorescence and real-time quantitative PCR methods. 【Results】 The PIZ/V5-BmIAP-dsRed vector effectively expressed BmIAP fusion protein of about 60 kD, and the mRNA expression level of BmIAP was up-regulated 45-fold in BmN-SWU1 cells of B.mori. After BmIAP was transiently expressed for 72 h using PIZ/V5-BmIAP-dsRed vector in the BmN-SWU1 cells, BmIAP significantly inhibited the apoptosis in the BmN-SWU1 cells at 12 h after induction by actinomycin D. 【Conclusion】 BmIAP can inhibit apoptosis in B.mori cells and is one of the inhibitor of apoptosis proteins in B.mori.

Key words: Bombyx mori; apoptosis; inhibitor of apoptosis protein(IAP); eukaryotic expression, transient expression