Composite

Part:BBa_M36014:Design

Designed by: Jesse Palmer, Elliot Lui   Group: Stanford BIOE44 - S11   (2011-05-02)

Glycogen Synthesis Actuator (alt2)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1996
    Illegal PstI site found at 2428
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1996
    Illegal PstI site found at 2428
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1996
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1996
    Illegal PstI site found at 2428
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1996
    Illegal PstI site found at 2428
    Illegal AgeI site found at 2620
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2344


Design Notes

Composite of genomic sequences (glgC and glgA) coding for proteins involved in e. coli glycogen biosynthesis. Using different RBSs so that secondary structures and self pairing don't occur. Since there are only two genes, strong 5'UTRs were available for both. Based on the literature, we predict this part would result in a lesser increase in glycogen production than either part BBa_M36012 or BBa_M36014 because the effect of over expressing glgA seems to have lesser influence than glgS. glgC coding sequence was retained in all three designs because the mutant used here resulted in increased glycogen production even with normal expression levels.

References

Buschiazzo A, et al. Crystal structure of glycogen synthase: Homologous enzymes catalyze glycogen synthesis and degradation. EMBO J. 2004;23(16):3196–3205.

Dedhia NN, Hottiger T, Bailey JE. 1994. Overproduction of glycogen in Escherichia coli blocked in the acetate pathway improves cell growth. Biotechnol Bioeng 44(1):132–139

Kumar, A., Ghosh, P., Lee, Y.M., Hill, M.A. and Preiss, J. (1989) Biosynthesis of bacterial glycogen: determination of the amino acid changes that alter the regulatory properties of a mutant E. coli ADP-glucose synthase. J. Biol. Chem. 264, 10 464–10 471.

Meyer CR, Ghosh P, Remy E, Preiss J. Cloning, expression, and nucleotide sequence of a mutant glgC gene from Escherichia coli B. J Bacteriol. 1992 Jul;174(13):4509–4512

Preiss J ADPglucose pyrophosphorylase: basic science and applications in biotechnology. Biotechnol Annu Rev. 1996; 2: 259–279.

Preiss J., Shen L., Greenberg E., Gentner N. Biosynthesis of bacterial glycogen. IV. Activation and inhibition of the adenosine diphosphate glucose pyrophosphorylase of Escherichia coli. Biochemistry. 1966;5:1833–45.