The Arabian camel Camelus dromedarius heat shock protein 90α: cDNA cloning, characterization and expression

Int J Biol Macromol. 2015 Nov;81:195-204. doi: 10.1016/j.ijbiomac.2015.07.058. Epub 2015 Jul 31.

Abstract

Heat shock protein 90 (Hsp90) is a highly conserved ubiquitous molecular chaperone contributing to assisting folding, maintenance of structural integrity and proper regulation of a subset of cytosolic proteins. In the present study, a heat shock protein 90α full length coding cDNA was isolated and cloned from the Arabian one-humped camel by reverse transcription polymerase chain reaction (RT-PCR). The full length cDNA sequence was submitted to NCBI GeneBank under the accession number KF612338. The sequence analysis of the Arabian camel Hsp90α cDNA showed 2202bp encoding a protein of 733 amino acids with estimated molecular mass of 84.827kDa and theoretical isoelectric point (pI) of 5.31. Blast search analysis revealed that the C. dromedarius Hsp90α shared high similarity with other known Hsp90α. Comparative analyses of camel Hsp90α protein sequence with other mammalian Hsp90s showed high identity (85-94%). Heterologous expression of camel Hsp90α cDNA in E. coli JM109 (DE3) gave a fusion protein band of 86.0kDa after induction with IPTG for 4h.

Keywords: Arabian camel; Camelus dromedarius; Chaperones; Heat shock protein; Reverse transcription polymerase chain reaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Camelus / classification
  • Camelus / genetics*
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Gene Order
  • Genetic Vectors / genetics
  • HSP90 Heat-Shock Proteins / chemistry
  • HSP90 Heat-Shock Proteins / genetics*
  • HSP90 Heat-Shock Proteins / metabolism
  • Male
  • Models, Molecular
  • Molecular Sequence Data
  • Phylogeny
  • Protein Conformation
  • Recombinant Fusion Proteins / genetics
  • Sequence Alignment

Substances

  • DNA, Complementary
  • HSP90 Heat-Shock Proteins
  • Recombinant Fusion Proteins