Project's information

Project's title Producing recombinant conotoxin and testing it’s analgesic activity
Research hosting institution Institute of Biotechnology
Project leader’s name As.Prof. Nguyen Bich Nhi
Project duration 01/01/2011 - 01/01/2012
Project’s budget 300.000.000 VND
Classify Fair
Goal and objectives of the project
  1. Cloning 1-2 genes encoding for conotoxins with high pharmaco-bioactive value from some special Conus collected from Nha Trang seacoach, Vietnam;
  2. Expression and purufication and characterization of recombinant conotoxins;
  3. Determination of analgesic activity of recombinant conotoxins on the animal experimental models.
Main results

Theoretical results

  • 2 conotoxins were cloned, sequenced from Conus species collected from Nha Trang seacoach, Vịetnam: (i) ω-conotoxin MVIIA (ω-CTX) with 75bp in size, encoding for 25 amino acids from Conus magus and (ii) µO-conotoxin MrVIB (µO-CTX) with 105 bp encoding for 31 amino acids from Conus marmoreus and expressed, purified 2 these conotoxins in fusion with thioredoxin in E. coli.
  • Protocol of expression and purification of the recombinant fusion protein Trx-ω-CTX (thioredoxin and ω-conotoxins) in E. coli with the yield 60 mg/L and basic properties of Trx-CTX was developed.
  • The analgesic activity of Trx-ω-CTX and Trx-µO-CTX was measured in hot-plate assay with mice of Swiss race weighing 20g. The results showed that Trx-CTX possess an analgesic activity better than Morphine hydrochloride.
  • LD50 of Trx-CTX is detected at the dose of 775 µg/g mouse body weight (130 -260 times higher than analgesis dose). The result showed the Trx-CTX no toxic on the experimental mice.
  • Studied on neuroprotective activity of Trx-CTX in mouse model of global cerebral ischemia and focal brain ischemia. The results showed that in global cerebral ischemia Trx-CTX has decreased neuronal injury and death of CA1 cells of dorsal hippogampus. In other model Trx-CTX also decreased infarct size of the ischemic brain.

Applied results: Trx-CTX proof to possess analgesic activity and neuroprotectiv effect and could be used in further researchs for medicinal purpose.
Novelty and actuality and scientific meaningfulness of the results
  • The recombinant ω-conotoxin MVIIA and µO-CTX were the first time produced and characterized in Vietnam.
  • Trx-CTX possess an analgesic activity better and activity time longer than morphine hydrochloride and its LD50 is detected at the dose of 775 µg Trx-CTX/g mouse body weight.
Products of the project

Scientific papers in referred journals (list): 4

  • Bui Thi Huyen, Doan Viet Binh, Nguyen Thi Kim Dung, Le Thi Bich Thao, Nguyen Bich Nhi, Phan Van Chi (2012) Research on neuroprotective effects of a fusion protein of ω-conotoxin MVIIA and thioredoxin. Journal of Biotechnology 34(4): 515-520.
  • Doan Viet Binh, Bui Thi Huyen, Nguyen Thi Kim Dung, Le Thi Bich Thao, Nguyen Bich Nhi, Phan Van Chi (2012) Research on acute toxicity and analgesic activity of a fusion protein of ω-conotoxin MVIIA and thioredoxin. Journal of Biotechnology 34(2): 241-245.
  • Le Thi Bich Thao, Bui Thi Huyen, Bui Quang Nghi, Doan Viet Binh, Nguyen Bich Nhi, and Phan Van Chi (2011) Study on the conopeptides from Cone snails collected from Nha Trang. Journal of Biotechnology 33(3): 74-80.
  • Le Thi Bich Thao, Bui Thi Huyen, Bui Quang Nghi, Doan Viet Binh, Nguyen Bich Nhi, and Phan Van Chi (2011) Study on the conopeptides from Cone snails collected from Nha Trang. Proceedings of the Workshop Coastal Marine Biodiversity and Bioresources of Vietnam and Adjacent areas to the South China Sea, Nha Trang, Vietnam, November 24-25, 2011, pp.114-118.

Technological products (describe in details: technical characteristics, place): Protocols of epression and purification of the recombinant fusion protein Trx-CTX  in E. coli was developed and purified Trx-CTX was stored in -20°C at Protein Biochemistry Lab, Institute of Biotechnology.

Other products (if applicable): 1 PhD thesis (Bui Thi Huyen, 2011-2015)