Nucleic acid construct for endogenous expression of RNA polymerase in cell

• Main Authors: SHIROTA JUN, XUE YINGE, CHAI ZHI, YU XUE, LIU SHUAILONG, GUO MIN, LI HAIYANG, WANG HAIPENG
• Format: Patent
• Language: Chinese; English
• Published: 05.12.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention provides a nucleic acid construct for endogenous expression of RNA polymerase in cells, and particularly discloses a novel nucleic acid construct capable of greatly enhancing protein translation efficiency, the nucleic acid construct provided by the invention is composed of coding sequences of promoters (such as RNR2, ADH1, GAPDH, TEF1, PGK1, SED1 and the like) with specific starting strength and RNAP (such as T7RNAP, T3RNAP, T4RNAP, T5RNAP and the like) proteins, and the coding sequences of the proteins are shown in the description. When the nucleic acid construct disclosed by the invention is applied to a yeast in-vitro protein synthesis system, the relative light unit value (RLU) of the activity of the synthesized luciferase is very high, and the same effect as that of exogenously added T7RNAP can be achieved and can reach 6.6 * 10 < 7 >. 本发明提供了一种细胞中内源性表达RNA聚合酶的核酸构建物，具体地，发现了一种可大幅度增强蛋白质翻译效率的新型核酸构建物，本发明的核酸构建物由具有特定启动强度的启动子(如RNR2、ADH1、GAPDH、TEF1、PGK1、SED1等)和RNAP(如T7RNAP、T3RNAP、T4RNAP、T5RNAP等)蛋白的编