Construction of human paraoxonase 3 gene expression vector
A paraoxonase and gene technology, applied in the field of biotechnology and pharmacy, can solve the problems of long-term multiple medication, slow onset of action, short action time, etc. Effect
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[0039] 1. The pMD18T-hPON3 plasmid was constructed and preserved by our laboratory. Using pMD18T-hPON3 as a template, primers 1 and 2 were used to amplify the full-length gene of hPON3 with a total of 1065 bp by PCR, and inserted into the corresponding multiple cloning site of the pCI plasmid after double digestion with EcoR I and SalI. The hPON3 gene is under the control of the CMV promoter. Sequencing proved that the result was completely consistent with the designed sequence (see Figure 1 for the process flow). Transform pCI-hPON3 with CaCl 2 The competent TOP10 Escherichia coli was prepared by the method, and the Escherichia coli was amplified according to the operation process of the third edition of "Molecular Cloning", and a large number of plasmids were extracted by the PEG method. The purity and yield of the extracted plasmids were determined by UV spectrophotometry, and the plasmid concentration was adjusted to 2 μg / μl with PBS. All biochemical reagents were purch...
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