BIOINFORMATICS, TISSUE DISTRIBUTION, AND SUBCELLULAR LOCALIZATION ANALYSES OF FK506 BINDING PROTEIN 12B FROM SILKWORMS

Arch Insect Biochem Physiol. 2016 Feb;91(2):109-23. doi: 10.1002/arch.21312. Epub 2015 Dec 17.

Abstract

FK506 binding proteins (FKBPs) are intracellular receptors of the immunosuppressant FK506 and play important roles in the correct folding of new proteins and the self-assembly of biological macromolecules. FKBP12 is a member of the FKBP family that is widely expressed and highly conserved in many species. In this study, we identified the complete cDNA sequence encoding the FKBP12 ortholog in Bombyx mori, named Bm-FKBP12B (GenBank accession no. DQ443423). Multiple-sequence alignment among different species revealed a high similarity among FKBP12 paralogs and orthologs. Bioinformatics analysis of the Bm-FKBP12B gene showed that it is located on chromosome 20 and consists of three exons and two introns. We cloned, expressed, and purified the Bm-FKBP12B protein in Escherichia coli and generated a specific polyclonal antibody against Bm-FKBP12B. The real-time quantitative reverse-transcription (qRT) PCR and Western blotting results showed that Bm-FKBP12B was present throughout all of the development stages, but it was abundant in the adult and embryo stages. Bm-FKBP12B expression was higher in the silk gland and gut, suggesting that it might play important roles in regulating gene expression in the silk gland and during silk fiber formation. Bm-FKBP12B protein was distributed in the cytoplasm, nucleus, and nuclear membrane.

Keywords: Bombyx mori; FKBP; FKBP12; expression.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bombyx / genetics
  • Bombyx / metabolism*
  • Computational Biology
  • Gene Expression
  • Insect Proteins / genetics
  • Insect Proteins / isolation & purification
  • Insect Proteins / metabolism
  • Molecular Sequence Data
  • Phylogeny*
  • Protein Conformation
  • RNA Interference
  • Sequence Homology, Amino Acid
  • Tacrolimus Binding Protein 1A / genetics
  • Tacrolimus Binding Protein 1A / isolation & purification
  • Tacrolimus Binding Protein 1A / metabolism*

Substances

  • Insect Proteins
  • Tacrolimus Binding Protein 1A