Transmembrane transduction method of low temperature lipase gene from marine microorganisms using ptd-tat

Abstract

The present invention is applicable to the technical field of fusion expression of transmembrane transduction gene fragments and lipase, and provides a transmembrane transduction method utilizing PTD-Tat to realize low-temperature lipase gene of marine microorganisms, comprising the following steps: 1. Design and obtain Lipase ‑tat fusion gene; 2. Construction of pET28a‑Lipase‑tat vector: First, digest Lipase‑tat with Nco I and Xho I, and recover the digested fragments from the gel; then use Nco I and Xho I to digest the pET28a vector, and recover from the gel Linearized pET28a vector fragment; then ligated the enzyme-digested fragment and the pET28a vector fragment to obtain the recombinant expression vector plasmid; 3. Prokaryotic expression and purification of Lipase-tat protein. Thereby, the present invention can realize the transmembrane transduction of the low-temperature lipase gene of marine microorganisms, so that the weight loss drug prepared from the recombinant protein can enter the cells; at the same time, it is the transmembrane transduction mechanism of the PTD-Tat gene and the low-temperature lipase gene of marine microorganisms It provides a theoretical and practical basis for its effective application in protein drug delivery.

Description

technical field [0001] The invention relates to the technical field of fusion expression of transmembrane transduction gene fragments and lipase, in particular to a method for transmembrane transduction of low-temperature lipase gene of marine microorganisms by using PTD-Tat. Background technique [0002] With the development of society and economy, obesity, as a multifactorial chronic metabolic disease, is increasing day by day, and has reached a historical peak in the past 20 years. At present, the wave of weight loss is sweeping across the world. Nowadays, almost all weight loss drugs use lipase inhibitors. Their working principle is to reduce the absorption of fat in substances by the human body by inhibiting the decomposition and catalysis of lipase in the intestinal tract on substances that have entered the human body but have not yet formed human body fat. Control and treat obesity. However, lipase inhibitors can only work in the intestinal tract and cannot enter hum...

AI Technical Summary

Problems solved by technology

[0005] If PTD-Tat is used to carry the low-temperature lipase gene of marine microorganisms to achieve transmembrane transduction, it can not only decompose the substances that have entered the human body but have not yet formed human fat in the intestinal tract, but also enter the deep tissues of the human body for drug action , this way of losing weight will be a brand-new idea of ​​losing weight, but there is no research report on the method of using PTD-Tat for lipase transmembrane transduction

[0006] In summary, there are obviously inconveniences and defects in the actual use of the existing technology, so it is necessary to improve

Images