Composite

Part:BBa_M36012:Design

Designed by: Elliott Lui and Jesse Palmer   Group: Stanford BIOE44 - S11   (2011-04-28)

Glycogen Synthesis Actuator (ideal)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1958
    Illegal PstI site found at 2390
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1958
    Illegal PstI site found at 2390
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1958
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1958
    Illegal PstI site found at 2390
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 618
    Illegal EcoRI site found at 1115
    Illegal EcoRI site found at 1958
    Illegal PstI site found at 2390
    Illegal AgeI site found at 2582
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2306


Design Notes

Composite of genomic sequences (glgC, glgA, and glgS) coding for proteins involved in e. coli glycogen biosynthesis. Using different RBSs so that secondary structures and self pairing don't occur. Only two strong UTRs were available, thus glgC and glgS coding sequences were paired with strong 5'UTRs and glgA with a medium strength 5'UTR. This particular arrangement was chosen because the effect of over expressing glgA seems to have lesser influence than glgC or glgS. We were unable to synthesize this part for experimentation due to size restrictions.

Source

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.