Construction method of humanized helicobacter pylori cagA eukaryotic expression vector

Abstract

The invention belongs to the field of biologic medicines, and particularly relates to a construction method of a humanized helicobacter pylori cagA eukaryotic expression vector. The construction method comprises a codon optimization step, in the codon optimization step, a humanized cagA gene is synthetized, and the nucleotide sequence of the humanized cagA gene is as shown in SEQID NO:1 in a sequence table. The humanized helicobacter pylori cagA eukaryotic expression vector constructed by the method can massively express cagA protein in gastric cancer cells, and conditions are created for researching the carcinogenic and pathogenic mechanism of protein coded by the cagA gene. The technical scheme of the construction method can be applied to practice of researching the pathogenic mechanismof helicobacter pylori.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for constructing a humanized Helicobacter pylori cagA eukaryotic expression vector. Background technique [0002] Helicobacter pylori (H. pylori) is a spiral-shaped Gram-negative microaerophilic bacterium that infects half of the world's population and is the main cause of gastritis, peptic ulcer, and gastric cancer. Among the infected people, most people cause asymptomatic gastritis, about 20-25% develop chronic gastritis, 10% develop peptic ulcer, and 1-3% develop gastric cancer. The occurrence of gastric cancer is the result of the comprehensive action of multiple factors, mainly including Helicobacter pylori infection, host genetic factors and environmental factors. Among them, Helicobacter pylori infection is the most important initiating factor of gastric cancer, and has been listed as a class I carcinogen of human gastric cancer by the World Health Organiza...

AI Technical Summary

Problems solved by technology

However, the above-mentioned technical scheme has the following problems: although the prokaryotic gene cagA is constructed into a eukaryotic expression vector and the eukaryotic expression of the prokaryotic gene is realized, the heterologous expression of the prokaryotic cagA gene in eukaryotic cells has a low expression level and a low expression level. Unstable, even when changing culture conditions or gastric cancer cell lines, gene expression often occurs, which cannot meet the requirements of further research on cagA regulatory pathways at this stage

Because the expression of cagA gene is too low or not expressed, the amount of related protein produced is less or no related protein is produced, and all cagA gene effector proteins, effector genes or downstream pathways cannot be activated completely, which makes the cagA gene cause gastric cancer. In-depth study of the molecular mechanism is limited

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