Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

General purpose method for preparing integration type cell immortalization vector

An immortalization and integrated technology, applied in the biological field, can solve the problems of low efficiency of immortalization and narrow host range of type II immortalization carriers, and achieve the effect of efficient immortalization transformation, wide host range and broad-spectrum transformation effect

Inactive Publication Date: 2008-07-09
OCEAN UNIV OF CHINA
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing a universal integrated cell immortalization carrier, to overcome the above-mentioned defects of the existing immortalization carrier, and to better solve the problem of low immortalization efficiency of class I immortalization carrier and class II immortalization The problem of the narrow host range of chemicalized vectors

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • General purpose method for preparing integration type cell immortalization vector
  • General purpose method for preparing integration type cell immortalization vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0011] Materials used in the present invention: plasmid pUC SV40-2895-1 containing SV40T gene (see Genbank accession no: AF316139, its nucleotide sequence is shown in SEQ ID NO.2 in the sequence listing, size 2.5kb) was purchased from American Type Culture Collection Center (ATCC), the SV40T gene contains small T antigen coding sequence (its nucleotide sequence is shown in SEQ ID NO.3 in the sequence listing); plasmids pLXRN (6.4kb), pVSV-G and GP- 293 cells were purchased from Clontech Company in the United States; NIH3T3 cells were donated by Professor Li Jing from the Institute of Marine Medicine, Ocean University of China; restriction enzymes BamH I, Xho I and LA Taq DNA polymerase were purchased from Dalian Bao Biological Company, and T4 DNA ligase was purchased from From Promega Company in the United States; DNA product purification kit and agarose gel DNA recovery kit were purchased from Beijing Tianwei Times Company; endotoxin-free plasmid extraction buffer was purchase...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a process for preparing a general integrated type cell immortalized vector. The process for preparation comprises following steps: firstly, directionally inserting a cell immortalized gene-simian virus 40 large T antigens (SV40T) gene with the broad spectrum transforming effect into a retroviral vector through a DNA recombinant technique to construct a recombinant expression vector, then, enabling the recombinant vector and packing plasmid pVSV-G to cotransfect GP-293 cells, and preparing recombinant pantropic reverse transcription virus particles which carry a SV40T gene, wherein the recombinant pantropic reverse transcription virus which is obtained is the general integrated type cell immortalized vector. The immortalized vector can enter into all species of animal cells and can integrate the carried SV40T gene to gene groups of target cells which is in mitotic phase to obtain inheritable and stable expression, and thereby integrated transformation with high efficiency of the target cells can be guaranteed. The invention can provide an effective means for transgenic immortalized establishment and studies of various animal cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for preparing a universal integrated cell immortalization carrier; more specifically, it relates to a method consisting of a simian virus 40 large T antigen (SV40T) gene and a pantropic retroviral vector. The preparation method of the recombinant virus. Background technique [0002] In life sciences and biotechnology, there is a broad demand for cell lines. The short-term culture of animal cells is relatively easy, but it is quite difficult to establish immortal cell lines. Immortalization genes are introduced into in vitro cultured cells to induce their ability to obtain unlimited proliferation, which is currently a commonly used method for line establishment. In this process, the vector carrying and expressing the immortalization gene is called the cell immortalization vector. Simian virus 40 large T antigen (SV40T) gene can induce the transformation of many types of cell...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/867C12N15/37C12N7/01
Inventor 胡国斌王丹宋珊珊王锡亮
Owner OCEAN UNIV OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com