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Molecular cloning and characterization of twist gene in Bombyx mori.

Authors
  • Guo, Min
  • Wang, Yong
  • Shi, Jing
  • Kang, Lequn
  • Yao, Qin
  • Wang, Fenghua
  • Qin, Lügao
  • Chen, Keping
Type
Published Article
Journal
Molecular and Cellular Biochemistry
Publisher
Springer-Verlag
Publication Date
Feb 01, 2011
Volume
348
Issue
1-2
Pages
69–76
Identifiers
DOI: 10.1007/s11010-010-0639-7
PMID: 21076855
Source
Medline
License
Unknown

Abstract

The twist genes are an evolutionarily conserved group of regulatory basic helix-loop-helix (bHLH) transcription factors. In present study, the twist gene was firstly cloned from Bombyx mori and was designated as BmTwist. Sequence analysis showed that BmTwist cDNA contains a 798 bp open reading frame, encoding a peptide of 266 amino acid residues. Sequence alignment showed that BmTwist protein shared extensive homology with other invertebrate Twist proteins in bHLH motif. RT-PCR and western blot analyses revealed that BmTwist expressed in all developmental stages of B. mori larvae various larval tissues. Here the authors also presented the results of prokaryotic expression, purification, and polyclonal antibody production of the BmTwist protein. Immunofluorescence of BmTwist in BmN cells using the antibodies showed that BmTwist protein was located in both the nucleus and cytoplasm. Furthermore, using B. mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system, the authors expressed a recombinant twist protein in BmN cell line. The obtained results, especially the preparation of polyclonal antibodies against BmTwist, will greatly facilitate further studies to explore biological functions of BmTwist protein such as identifying its potential binding partners.

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