Investigating protein function in biomineralized tissues using molecular biology techniques

Methods Enzymol. 2013:532:367-88. doi: 10.1016/B978-0-12-416617-2.00017-5.

Abstract

We describe modern molecular biology methods currently used in the study of biomineralization. We focus our descriptions on two areas of biomineralization research in which these methods have been particularly powerful. The first area is the use of modern molecular methods to identify and characterize the so-called occluded matrix proteins present in mineralized tissues. More specifically, we describe the use of RNA-seq and the next generation of DNA sequencers and the use of direct protein sequencing and mass spectrometers as ways of identifying proteins present in mineralized tissues. The second area is the use of molecular methods to examine the function of proteins in biomineralization. RNA interference (RNAi), morpholino antisense, and other methods are described and discussed as ways of elucidating protein function.

Keywords: DNA sequencing; Mass spectrometry; Morpholino antisense; Proteomics; RNA interference; RNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calcification, Physiologic
  • Gene Knockdown Techniques*
  • Humans
  • Molecular Biology
  • Morpholinos / genetics
  • Proteins / genetics
  • Proteins / metabolism
  • RNA Interference
  • Sequence Analysis, DNA

Substances

  • Morpholinos
  • Proteins