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

Construction of whole genome high-throughput cloning vector

A whole genome and cloning carrier technology, applied in the field of biomedicine, can solve the problems of difficult whole genome industrial amplification, difficult culture of fusion cells, low cell fusion rate, etc., to increase the success rate and fusion rate, easy to subculture and The effects of industrial expansion and less death

Active Publication Date: 2019-06-11
翁炳焕
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to solve the problems of low cell fusion rate, poor screening method, difficult cultivation of fused cells, and difficulty in the industrial amplification of the whole genome, the inventors proposed the present invention

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
  • Construction of whole genome high-throughput cloning vector
  • Construction of whole genome high-throughput cloning vector
  • Construction of whole genome high-throughput cloning vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0026] Combine below figure 1 , figure 2 , image 3 and Figure 4 , the embodiments of the present invention are described in detail.

[0027] 1. Construction of retroviral recombinant vectors pLPCX-hTERT and pLXN-SV40LT

[0028] Subcloned hTERT from PIRES2-EGFP-hTERT to the EcoRI site of pLPCX to construct the retroviral recombinant vector pLPCX-hTERT; subcloned SV40LT from pMFGSV40tsLT to the EcoRI / BamHI site of pLXN to construct the retroviral recombinant vector pLXN -SV40LT.

[0029] Reagents: pLPCX, pLXSN retroviral vectors are products of Clontech, USA; PIRES2-EGFP-hTERT or pCIneo-hTERT or, SV40DNA (strain 776) are products of Invitrogen, USA; PCR amplification kit and calcium phosphate precipitation transfection kit Purchased from Invitrogen Company in the United States; E.coliDH5-alpha Escherichia coli was preserved as the unit; Endotoxin-free plasmid purification kit was purchased from QIGEN Company in Germany; EcoR I, Not I, BamH I, and endonucleases were from ...

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 construction of a whole genome high-throughput cloning vector in the field of biomedicine. The construction is characterized by comprising the steps of constructing a reverse transcription virus vector containing hTERT and SV40LT; conducting transfecting on PT67 packaging cells; using g418 and PM for combined screening to obtain plasmid packaging PT67 cells; extracting a culture solution of the plasmid packaging PT67 cells, and conducting transfecting on sp2 / 0 cells; carrying out combined screening by using G418 and PM to obtain highly immortalized plasmid packaging sp2 / 0 cells; after an exogenous whole genome is implanted, using HAT, G418 and PM for combined screening to obtain a highly immortalized hybrid strain capable of being cryopreserved and subjected to whole genome amplification. The hybrid strain can be used for industrial preparation of rare disease whole genome and construction of an immortalized gene bank of the rare disease whole genome.

Description

technical field [0001] The invention relates to the construction of a whole-genome high-throughput cloning vector in the field of biomedicine, which is mainly used for the collection and industrial preparation of the whole genome of rare diseases, and provides renewable whole-genome resources for clinical, scientific research, pharmaceutical and diagnosis. Background technique [0002] In order to better carry out research on pathogenesis, clinical treatment and experimental diagnosis, more and more domestic and foreign institutions have focused on the collection of various case samples and the construction of sample banks. [0003] Various types of cell banks have been established in the United States, the United Kingdom, Japan and China, such as the American Standard Cell Bank (ATCC), the Human Genetic Mutation Cell Bank (HGMR), and the Cell Aging Cell Bank (CAR); the British Embryonic Stem Cell Bank Japanese deciduous tooth stem cell research bank; China's important medic...

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): C12N15/867C12N15/65C12N5/10
Inventor 翁炳焕黄荷凤马端
Owner 翁炳焕
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