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Recombinant lentiviral vector and preparation method and application thereof

A technology of recombinant lentivirus and lentiviral vector, which is applied in the biological field, can solve the problem that it cannot be used to study the transcriptional activity of regulatory elements, and achieves the effects of easy industrial production, low toxicity and simple preparation method.

Inactive Publication Date: 2015-06-17
SHENZHEN INST OF ADVANCED TECH
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AI Technical Summary

Problems solved by technology

Therefore, this expression vector cannot be used to study the functions of regulatory elements such as transcriptional activity of promoters and / or enhancers

Method used

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  • Recombinant lentiviral vector and preparation method and application thereof
  • Recombinant lentiviral vector and preparation method and application thereof
  • Recombinant lentiviral vector and preparation method and application thereof

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Embodiment Construction

[0050] The following description is a preferred embodiment of the present invention, it should be pointed out that for those skilled in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications are also considered Be the protection scope of the present invention.

[0051] The methods used in the following examples are conventional methods unless otherwise specified, and the specific steps can be found in: "Molecular Cloning: A Laboratory Manual" (Sambrook, J., Russell, David W., Molecular Cloning: A Laboratory Manual, 3 rd edition, 2001, NY, Cold Spring Harbor). The primers and DNA sequences used were synthesized by Shanghai Invitrogen Company.

[0052] The pLVX-IRES-ZsGreen1 lentiviral vector used in the examples of the present invention was purchased from Clontech, the strain containing the pGL4.10[luc2] plasmid was purchased from Promega, the DH5α used was commerci...

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Abstract

The present invention provides a lentiviral vector, and the lentiviral vector is obtained by inserting firefly luciferase coding gene sequence in the 2180bp of pLVX-IRES-ZsGreen1 lentiviral vector. The invention also relates to a host cell containing the lentiviral vector, a construction method of the lentiviral vector, and application of the lentiviral vector in analysis of function or regulation effect of mammalian cell promoters and / or enhancers. The lentiviral vector provides a fast, efficient, intuitive, simple and easy method for finding, analysis and application of target protein expression or expression level regulation promoters and / or enhancers.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant lentiviral vector and its preparation method and application. Background technique [0002] Plasmid-based transcriptional reporter vectors commonly used today can interfere with the studied reporter network due to their extrachromosomal episomal properties. Lentiviral-based reporter vectors can be integrated into cell chromosomes, providing a more accurate model for studying transcriptional activation and epigenetic effects. [0003] Clontech's pLVX-DD-ZsGreen1 lentiviral reporter vector is a promoter-less reporter vector, and its reporter gene is green fluorescent protein DD-ZsGreen1, which can effectively transduce dividing and non-dividing mammalian cells for detection of different Transcriptional activity of the promoter. However, the vector used to screen for stably transfected cell lines is puromycin, which is highly toxic to cells and easily causes cell death. ...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10C12N15/66C12Q1/68
Inventor 万晓春阮庆国王绍文
Owner SHENZHEN INST OF ADVANCED TECH
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