Method for efficiently expressing panda DNAJC12 gene
By optimizing the sequence of the giant panda DNAJC12 gene and adding His-tag, a prokaryotic expression system was constructed, which solved the problem of expression and purification of the giant panda DNAJC12 gene, and achieved efficient and efficient expression and purification methods.
CN119709805APending Publication Date: 2025-03-28SICHUAN RES INST OF GIANT PANDA SCI
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Patent Information
- Application Number
- CN202411903528.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-23
- Publication Date
- 2025-03-28
AI Technical Summary
Technical Problem
The prior art is difficult to efficiently express and isolate the recombinant protein of the giant panda DNA JC12 gene, and the prokaryotic cell expression system has not been established, resulting in limited research and application.
Method used
By codon optimization of the giant panda DNA JC12 gene sequence and adding His-tag sequence to the C-terminus, a prokaryotic expression system was constructed, and the pET-30a(+) vector and BL21(DE3) strain were used for expression and purification.
Benefits of technology
The efficient expression and simplified purification of the giant panda DNA JC12 gene were achieved, reducing research costs, overcoming sample source limitations, and improving experimental efficiency.
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Abstract
The invention discloses a method for efficiently expressing a panda DNAJC12 gene, and relates to the technical field of gene engineering. The method comprises the following steps: carrying out codon optimization on a panda DNAJC12 gene with a nucleotide sequence as shown in SEQ ID NO.1 to obtain an optimized sequence with a nucleotide sequence as shown in SEQ ID NO.2, sequentially adding an enzyme digestion cloning site and His-tag to the C-terminal of the obtained optimized sequence, connecting the obtained optimized sequence to a prokaryotic plasmid, and transferring the prokaryotic plasmid into a prokaryotic expression bacterium to obtain a recombinant expression bacterium; and culturing the obtained recombinant expression bacteria, and carrying out Ni-column purification to obtain the panda DNAJC12 protein. According to the method provided by the invention, the panda DNAJC12 gene can be efficiently expressed without adding an inducer, and a basis is provided for further researching the structure function and application of the DNAJC12.
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