Application of tRNA subjected to anticodon editing as molecular switch in precise control of protein expression and virus replication
An anti-codon, protein expression technology, applied in the direction of DNA / RNA fragment, virus / phage, recombinant DNA technology, etc., to achieve good application prospects and improve the effect of safety
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[0057] method
[0058] 1. Materials and methods
[0059] 1.1 Samples, cells and virus strains
[0060] Human embryonic kidney cells (HEK293T) were preserved by our laboratory. The pDC-315-GFP plasmid (using the pEGFP-N1 plasmid as a template, PCR amplified EGFP gene, and cloned it into the pDC-315 vector to obtain pDC-315-GFP) was constructed and preserved by our laboratory. Lentiviral packaging system plasmids PLVX-IRES-ZSGreen1, HIV-1-Luc (use pGL3-Basic vector as template, PCR amplify luciferase gene, clone it into pFUGW vector to obtain HIV-1-Luc), psPAX2 and pMD2.G Plasmids are kept by our laboratory. The pUC57 plasmid was purchased from Beijing Qingke Biotechnology Co., Ltd. DH5α competent cells used for plasmid transformation were purchased from Beijing Qingke Biotechnology Co., Ltd. (Beijing, China). Three anticodon-edited tRNAs——Arg UGA tRNA, Gly UGA tRNA and Trp UGA The tRNAs are respectively obtained by transcribing the nucleotide sequences shown in SEQ ID N...
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