African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12182

Full Length Research Paper

Expression and functional analysis of apoptosis-related gene BmICAD in silkworm, Bombyx mori

Wei Yu*, Yue Ma and Yao-Zhou Zhang
Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Email: [email protected]

  •  Accepted: 27 July 2011
  •  Published: 19 October 2011

Abstract

The ICAD gene plays an important role in the growth and development processesin insects. We conducted a molecular cloning and functional analysis to study a specific silkworm gene BmICAD related to apoptosis. The BmICAD gene was obtained from the fifth instar larvae of the silkworm by RT-PCR and over-expressed in Escherichia coli as His-tagged fusion proteins. Subcellular localization of the protein indicated that BmICAD was found in the cytoplasm near the nucleus. RNAi assay indicated that the apoptosis rate of Bm5 cells increased markedly. A His pull-down assay was used to investigate proteins that bind to rBmICAD. Two proteins, DNA supercoiling factor (SCF) and secreted protein acidic and rich in cysteine (SPARC), were found. These results indicate that down regulation of BmICAD can increase the apoptosis rate of Bm5 cells, and that SCF and SPARC may have important roles by interacting with BmICAD during this process.

 

Key words: Bombyx mori, BmICAD, subcellular localization, RNAi, protein interaction.

Abbreviation

OFR, Open reading frame; CIDE, cell death-inducing DFF45-1ike effectorsCAD, caspase-activated DnaseICAD, inhibitor of caspase-3-activated DNAaseDFF, DNA fragmentation factorPCD, programmed cell deathSCF,supercoiling factorSPARC, secreted protein acidic and rich in cysteine.

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