Part:BBa_K4862001
NS3-ZIKA Virus
Basic Part: BBa_K4862001 (NS3-ZIKA Virus)
Profile
- Name: NS3-ZIKA Virus
- Base Pairs: 1857 bp
- Origin: Dengue virus 2, Synthetic
- Properties: Zika virus (ZIKV) infection is associated with severe neurological disorders and congenital malformation.
Usage and Biology
Zika virus (ZIKV) infection is associated with severe neurological disorders and congenital malformation. Despite efforts to eradicate the disease, there is neither a vaccine nor approved drugs to treat ZIKV infection. The NS2B-NS3 protease is a validated drug target since it is essential to polyprotein virus maturation [1].
RNA helicase is responsible for almost all metabolism of RNA; in the Dengue virus, NS3 protein contains the activity of multiple enzymes, including helicase. Inserting the NS3 sequence is for expressing the protein we need and testing whether the medicine finally injected would be effective. The other structure of the pET28a plasmid would not be changed.
While these two plasmids are constructed, they would be transplanted into BL21 bacteria, which can express the NS3 protein we need. After growing for a few hours, the solution (these two plasmids and the liquid medium) would be ultrasonically crushed, destroying the bacteria's basic structure. The answer, consisting of the proteins and bacteria fragments, would be purified for extracting the NS3 proteins.
The NS3 proteins have the ability as a Helicase, which would activate the helicase to work. The protein would be added to a fluorescence DNA vector to observe whether the helicase works. If so, the fluorescence would disappear; if not, the fluorescence would be observed, which also means the medicine for restraining the replication of DNA is working. As NS3 proteins are the main protein active helicase in the Dengue virus, if the ability is restricted, the duplication of the RNA of the bacteria would be stopped, which means the spread of the virus would be halted.
References
- Andrade MA, Mottin M, Sousa BKP, Barbosa JARG, Dos Santos Azevedo C, Lasse Silva C, Gonçalves de Andrade M, Motta FN, Maulay-Bailly C, Amand S, Santana JM, Horta Andrade C, Grellier P, Bastos IMD. Identification of novel Zika virus NS3 protease inhibitors with different inhibition modes by integrative experimental and computational approaches. Biochimie. 2023 Sep;212:143-152. doi: 10.1016/j.biochi.2023.04.004. Epub 2023 Apr 22. PMID: 37088408.
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