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Derivational expression method of glucose polycytidylate enzyme in expression system of lactobacillus casei

A technology of glucuronidase and Lactobacillus casei, applied in bacteria, hydrolase and other directions, can solve the problem of low efficiency

Inactive Publication Date: 2007-10-10
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Lactobacillus expression vectors have been successfully used to express different foreign proteins, but almost all of them are not efficient in expressing proteins using Lactobacillus as a host strain. Therefore, improving the ability of Lactobacillus to express foreign proteins has become the focus of research

Method used

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  • Derivational expression method of glucose polycytidylate enzyme in expression system of lactobacillus casei
  • Derivational expression method of glucose polycytidylate enzyme in expression system of lactobacillus casei
  • Derivational expression method of glucose polycytidylate enzyme in expression system of lactobacillus casei

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Embodiment Construction

[0063] The following examples describe the present invention in more detail:

[0064] 1 Xylose induces the expression of gusA

[0065] Using D-xylose as an inducer, pick pPG611.1 / L..casei 393 and pPG612.1 / L..casei 393, and inoculate them in MRS basic medium containing 2% xylose. Cultivate L.casei 393 without plasmid and pPG611.1 / L..casei 393 grown in 2% glucose as the control culture, and induce by static culture at 37°C overnight. .

[0066] 2 Determination of optimal xylose concentration

[0067] Inoculate an appropriate amount of pPG611.1 / L..casei 393 in MRS liquid medium containing different concentrations of xylose, and culture overnight at 37°C. The concentrations of D-xylose are 0.05%, 0.1%, 0.5%, 1 %, 2%, 3%, 4%, 5%, after the end of the induction, take the bacterial solution for electrophoresis analysis, and further conduct a preliminary quantitative analysis of the expression product by optical density scanning to obtain the optimal induction concentration.

[00...

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Abstract

This invention provides a method for induced expression of beta-glucosidase in Lactobacillus casei 393. The method comprises: expressing beta-glucosidase in Lactobacillus casei 393 with surface-expression vector pPG611.1 and secretive expression vector pPG612.1. The target protein is generated in host cells. The expressed proteins are combined with cell walls, and do not exist in the supernatant. This invention also optimizes the induction conditions. The expression system can be used to study antigen presentation.

Description

1. Technical field [0001] What the present invention relates to is a kind of preparation method of biological material, specifically 2. Background technology [0002] In recent years, people have been working on new oral vaccines to avoid the disadvantages of traditional injectable vaccines. A large number of bacteria, viruses and parasitic pathogens enter the body through the mucosal surface, so oral vaccines as a way of immunity are better than traditional injection vaccines; although injection vaccines can cause systemic immune responses, they cannot effectively induce the body to produce Secreted antibody sIgA; oral vaccines are not only convenient to use and low in cost, but for many pathogens, oral immunization may produce a herd immunity effect, that is, when only a few members of the group are immune, this immunity will be in the group These advantages apply especially to less industrialized countries. Therefore, the research and selection of live vectors for oral ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N9/24
Inventor 李一经唐丽杰葛俊伟任晓峰夏春丽
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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