Detection method of hereditary stability of gene vector for encoding muramidase-released protein and and application

A technology of genetic stability and gene carrier, applied in the field of detection of genetic stability of gene carrier, to achieve the effect of important application value

Inactive Publication Date: 2018-05-29
HANBANG MEDICAL SCI & TECH HARBIN CITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Few studies have been conducted on the stability of artificially preserved gene vectors encoding lysozyme-releasing proteins

Method used

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  • Detection method of hereditary stability of gene vector for encoding muramidase-released protein and and application
  • Detection method of hereditary stability of gene vector for encoding muramidase-released protein and and application
  • Detection method of hereditary stability of gene vector for encoding muramidase-released protein and and application

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Experimental program
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Embodiment 1

[0055] This embodiment provides a method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein, comprising transforming the gene carrier plasmid encoding a lysozyme-releasing protein into a host bacterial cell, and then converting the gene sequence containing the encoding lysozyme-releasing protein The host bacterium of the carrier plasmid is made into glycerol bacterium, and the genetic stability of the subcultured glycerol bacterium is identified by culturing and subculture of the glycerol bacterium.

[0056] In this example, DH5α competent cells were selected as E. coli competent cells. Of course, in other embodiments, other E. coli competent cells, such as TOP10 cells or BL21(DE3) cells, could also be selected for testing.

[0057] The specific steps of plasmid transformation are as follows:

[0058] 1.1 Take 100 μL of competent cells of Escherichia coli DH5α and add them to EP tubes, and thaw on ice;

[0059] 1.2 Take 1 μL of gene ca...

Embodiment 2

[0073] The present embodiment provides a method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein, and the method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein comprises transforming the viral plasmid of the gene carrier encoding a lysozyme-releasing protein into In the host bacteria cells, the host bacteria containing the virus plasmid of the gene carrier encoding the lysozyme releasing protein are made into glycerolbacteria, and the genetic stability of the passaged glycerolbacteria is identified by culturing and subculture of the glycerolbacteria.

[0074] In this example, TOP10 competent cells were selected as E. coli competent cells. Of course, in other embodiments, other E. coli competent cells, such as DH5α cells or BL21(DE3) cells, could also be selected for testing.

[0075] The specific steps of plasmid transformation are as follows:

[0076] 1.1 Add 100 μL of competent cells of E...

Embodiment 3

[0091] The present embodiment provides a method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein, and the method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein comprises transforming the viral plasmid of the gene carrier encoding a lysozyme-releasing protein into In the host bacteria cells, the host bacteria containing the virus plasmid of the gene carrier encoding the lysozyme releasing protein are made into glycerolbacteria, and the genetic stability of the passaged glycerolbacteria is identified by culturing and subculture of the glycerolbacteria.

[0092] In this example, TOP10 competent cells were selected as E. coli competent cells. Of course, in other embodiments, other E. coli competent cells, such as DH5α cells or BL21(DE3) cells, could also be selected for testing.

[0093] The specific steps of plasmid transformation are as follows:

[0094] 1.1 Add 100 μL of competent cells of E...

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Abstract

The invention provides a detection method of hereditary stability of gene vector for encoding muramidase-released protein and application, and belongs to the field of biological engineering. The detection method provided by the invention is characterized in that a host bacteria bacterial fluid containing gene vector plasmids for encoding muramidase-released protein is subjected to recovery culture, and is subjected to continuous multi-generation subculture as well as is sampled and detected at a certain generation interval, and the hereditary stability of the artificially preserved host bacteria containing the gene vector plasmids encoding muramidase-released protein is detected, thereby providing reliable support and guarantee for researching the gene vector for encoding muramidase-released protein. The detection method can be applied to the culture of the gene vector for encoding muramidase-released protein so as to provide reference for the research and the culture of the gene vector for encoding muramidase-released protein, and has important application values.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for detecting the genetic stability of a gene carrier encoding a lysozyme-releasing protein and its application. Background technique [0002] Streptococcus suis is a Gram-positive coccus with an capsule. It can be roughly classified as Lancefield group D (Lancefieldgroup D) group D streptococcus according to its cell wall antigenic composition. [0003] Streptococcus suis is mainly manifested as septicemia and focal lymph node suppurative disease in pigs. The natural infection sites of Streptococcus suis are the upper respiratory tract (especially tonsils and nasal cavity), reproductive tract and digestive tract of pigs. Pigs have a higher susceptibility among animals. Pigs of all ages can suffer from the disease, but the septicemia type and meningoencephalitis type are more common in piglets, and the suppurative lymphadenitis type is more common in middle-aged pigs. S...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12N15/31C12R1/19
CPCC07K14/315C12N15/70
Inventor 余美伦孟庆雪秦香芹李影韩伟丽何佳王春东苑玉凤王宁唐世梅
Owner HANBANG MEDICAL SCI & TECH HARBIN CITY
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