Difference between revisions of "Team:DUT China B"

description

• Design of DIP micro system

  Through the review of the related properties of microalgae, we learn the advantages of Chlamydomonas reinhardtii in photoautotrophy, secretion, and movement over traditional chassis organisms, and we also learn that in the field of chassis microorganism, related development is incomplete. As above two reasons, we want to use the C. reinhardtii to construct a “Real-time detection、Induced expression-separation and Purification”(DIP )micro-system, and we also want to transform C. reinhardtii into a chassis organism, to make more contributions to the development of synthetic biology research tools.

  This device in the left reflects our initial vision for a separation and purification system. Through the regulation of the upper valve, we can supplement the inorganic nutrients and water in the algae culture area; by adjusting the intermediate valve, we can realize functions of cleaning, adsorbing, washing, eluting before separation, and finally keep the eluent in the bottom tank for storage.

  Our microsystem consists of three parts: one is real-time monitoring, the other is to induce expression, and the third is to achieve separation and purification. We construct different fluorescent response monomers in fluorescent fiber hydrogels to detect different substances[2]; We design the fluorescent signal of hydrogel fibers that algae can respond to and express the target protein. The target protein is secreted into the surrounding liquid environment and enters the separation and purification system. After absorbed by the gel beads, washed and eluted in the micro device, we can obtain the purified protein and store them in a storage tank [3]. Based on the construction of the D-I-P micro-system, we intend to construct a stable light-inducing system in Chlamydomonas. Hopefully, we could make some contributions to synthetic biology.

• Advantages of the D-I-P system

  This system has following advantages:

  1. Chlamydomonas reinhardtii is a photosynthetic autotrophic organism, avoiding the issues of continuously adding organic carbon sources. It can continuously secreting proteins in the presence of light and inorganic nutrients;
  2. The micro-separation and purification devices simplifies the complicated process of traditional protein purification, spares the trouble of looking for the equipment required for the traditional purification method and the storage conditions specific for the purified protein, and reduces the cost of protein separation and purification;
  3. Our system can detect different substances and realize the real-time detection function and give feedback simultaneously as conditions in different scenarios changes .

inspiration

As a photosynthetic autotrophic microorganism, C. reinhardtii is a new synthetic biology chassis organism with great potential, which has aroused our interest greatly. From the research of the related aspects on the synthetic biology of Chlamydomonas, we know that there is little content about the induction system in the synthetic biology research of Chlamydomonas, while the induction system is essential for the construction of logic gates in synthetic biology. Therefore, we hope to construct a light-induced system in Chlamydomonas. In the search for practical applications of light-inducing systems, we have noticed that the secretory system is one of the most studied problems in Chlamydomonas. Inspired by our test project (DLUT_CHINA_B) [1] in the last year, we naturally thought of to construct a system that can detect, secret and give feedbacks at the same time, so our system prototype was born

[1] CROZET P, NAVARRO F J, WILLMUND F, et al. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii [J]. ACS Synthetic Biology, 2018, 7(9): 2074-86.

[2] Francisco J. Navarro, David C. Baulcombe. miRNA-Mediated Regulation of Synthetic Gene Circuits in the Green Alga Chlamydomonas reinhardtii. ACS Synthetic Biology 2019, 8 (2) , 358-370. DOI: 10.1021/acssynbio.8b00393.

[3] MURPHY T W, SHENG J, NALER L B, et al. On-chip manufacturing of synthetic proteins for point-of-care therapeutics [J]. Microsystems & Nanoengineering, 2019, 5(1): 13.

[4] https://2018.igem.org/Team:DLUT_China_B