mohammad tabish ahmed - eTheses Repository - University of ...

Chapter 6
In this chapter I will be looking to further characterise the chaperonins of Mycobacterium
tuberculosis in three different sets of experiments. For this reason, the chapter will be split
into 3 separate sections so that their background information and results can be discussed
separately. The three sections that will be discussed here are as follows:
6.1- Testing the ability of M. tuberculosis Cpn60.1 to function in M. smegmatis
6.2- Testing the existence of double ring M. tuberculosis Cpn60.2 by attempting to make
single ring mutants
6.3- Testing the hypothesis that phosphorylation is needed for improved oligomerisation
In section 6.1, since I will be referring to M. tuberculosis, M. smegmatis and E. coli
chaperonins, the prefixes Mtb, Ms, or Ec will be used. So, MtbCpn60.1 denotes M.
tuberculosis Cpn60.1, while MsCpn60.1 denotes M. smegmatis Cpn60.1, and EcGroEL
denotes E. coli GroEL. In all cases, unless mentioned otherwise, the cochaperonin in the
experiments was M. smegmatis Cpn10. To make it easier to follow, each strain will be
denoted with a letter:
Strain A- M. smegmatis mc 2 155
Strain B- M. smegmatis Δcpn60.1
Strain C- Strain B with pMsGroEL1 plasmid (Mscpn10-Mscpn60.1)
Strain D- Strain B with pMs10-MsCpn60.2 plasmid
Strain E- Strain B with pMs10-MsCpn60.3 plasmid
Strain F- Strain B with pMs10-EcGroEL plasmid
Strain G- Strain B with pMs10-MtbCpn60.1plasmid
Note: The constructs in strains D, E, and F were made by Tara Rao (PhD thesis, 2010,
University of Birmingham).
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