Difference between revisions of "Part:BBa K1021017"

Revision as of 02:32, 29 October 2013

PT7+crtB

This part is the crtB gene (the second component of the carotenoid biosynthesis pathway) downstream of the T7 promoter.

Adapted from KEGG reference pathways

crtB was originally from the 2008 Edinburgh team that isolated the coding sequence of crtB from Pantoea ananatis. crtB encodes phytoene synthase, part of the carotenoid biosynthesis pathway, which converts geranylgeranyl diphosphate to phytoene (Misawa et al., 1990).

The T7 promoter system was used in order to assemble multiple genes into a single vector. Fungal promoters are quite long and therefore difficult to assemble, and E. coli will often recombine over and remove direct repeats within DNA sequences. Because the T7 promoter is a short sequence, each of the carotenoid genes could be put behind the T7 promoter and assembled together for maximum simultaneous transcription of the carotenoid genes.

1. Misawa, N., Nakagawa, N., Kobayashi, K., Yamano, S., Nakamura, K., and Harashima, K. 1990. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. Journal of Bacteriology 172, 6704-6712.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 616
Illegal NgoMIV site found at 746
1000
COMPATIBLE WITH RFC[1000]

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 Adapted from KEGG reference pathways</center>
-crtB was originally from the 2008 Edinburgh team that isolated the coding sequence of  ''crtB'' from ''Pantoea ananatis.'' crtB encodes phytoene synthase, part of the carotenoid biosynthesis pathway, which converts geranylgeranyl diphosphate to phytoene (Misawa, N., Nakagawa, N., Kobayashi, K., Yamano, S., Nakamura, K., and Harashima, K. 1990. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. Journal of Bacteriology 172, 6704-612).
+crtB was originally from the 2008 Edinburgh team that isolated the coding sequence of  ''crtB'' from ''Pantoea ananatis.'' crtB encodes phytoene synthase, part of the carotenoid biosynthesis pathway, which converts geranylgeranyl diphosphate to phytoene (Misawa et al., 1990).
-The T7 promoter system was used in order to express multiple genes simultaneously. Fungal promoters are quite long, and prokaryotes have systems to splice out long repetitive sequences of DNA. Because the T7 is a short sequence, each of the carotenoid genes could be put behind the T7 promoter for maximum simultaneous transcription of the carotenoid genes.
+The T7 promoter system was used in order to assemble multiple genes into a single vector. Fungal promoters are quite long and therefore difficult to assemble, and ''E. coli'' will often recombine over and remove direct repeats within DNA sequences. Because the T7 promoter is a short sequence, each of the carotenoid genes could be put behind the T7 promoter and assembled together for maximum simultaneous transcription of the carotenoid genes.
+. Misawa, N., Nakagawa, N., Kobayashi, K., Yamano, S., Nakamura, K., and Harashima, K. 1990. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. Journal of Bacteriology 172, 6704-6712.
 <!-- Add more about the biology of this part here