Composite

Part:BBa_K3458004:Design

Designed by: Jiahua Zou   Group: iGEM20_GDSYZX   (2020-08-16)


GluD-1 Promoter + HQT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 225


Design Notes

The goal of our team is to specifically express HQT, a key enzyme for chlorogenic acid synthesis in the endosperm of rice. The promoter GluD-1 cannot drive the HQT express in Oryza sativa L. protoplasts. In order to test our expression system in protoplasts, we designed a 35s promoter + HQT (BBa_K3458004) composite element for comparison with this part.

Source

  • The GluD-1 promoter BBa_K3458002) is derived from the rice genome and has a full length of 1.6 kb. Since studies have shown that the 0.2kb promoter can specifically express foreign genes in rice endosperm, and the 1.2kb efficiency is the highest, our team chose the 1.2kb GluD-1 promoter. In order to meet the assembly requirements of biobrick, our team checked the predicted cis-elements and modified the illegal restriction sites of non-critical parts.
  • The HQT gene BBa_K3458001) we used comes from the genome of Lonicera japonica (GeneBank: JF261014.1). We removed the restriction site incompatible with the biobrick assembly standard and optimized the codon according to the codon preference of Oryza sativa L..The HQT was synthesized in TIANYI HUIYUAN Company.

References

[1]Peng X, Li W, Wang W, et al. Cloning and characterization of a cDNA coding a hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase involved in chlorogenic acid biosynthesis in Lonicera japonica[J]. Planta Med., 2010,76(16):1921-1926.

[2]Kawakatsu T, Yamamoto M P, Hirose S, et al. Characterization of a new rice glutelin gene GluD-1 expressed in the starchy endosperm[J]. J. Exp. Bot., 2008,59(15):4233-4245.

[3]Ye X, Al-Babili S, Kloti A, Zhang J, Lucca P, Beyer P, Potrykus I. 2000. Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287, 303–305.

[4]Paine JA, Shipton CA, Chaggar S, et al. 2005. Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nature Biotechnolgy 23, 482–487.

[5]Qu LQ, Takaiwa F. 2004. Evaluation of tissue specifificity andexpression strength of rice seed component gene promoters intransgenic rice. Plant Biotechnology Journal 2, 113–125.