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A549 nude mouse model of stably expressed luciferase and building and application thereof

A luciferase, stable expression technology, applied in applications, preparations for in vivo experiments, and cells modified by introducing foreign genetic material, etc., can solve the problems of large influencing factors, complicated operation, inconvenient research, etc., and achieve reliable results. , observe the effect of intuitive and convenient, good application prospects

Inactive Publication Date: 2013-02-20
SUZHOU RES INST OF TONGJI UNIV
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  • Abstract
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Problems solved by technology

However, most of the current transplanted lung cancer models need to be evaluated by anatomical methods to evaluate the efficacy of drugs. The operation is complicated, and the factors affected by human operation are large, which is not convenient for preclinical research.

Method used

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  • A549 nude mouse model of stably expressed luciferase and building and application thereof
  • A549 nude mouse model of stably expressed luciferase and building and application thereof
  • A549 nude mouse model of stably expressed luciferase and building and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Step 1. Cloning of luciferase gene and construction of lentiviral vector:

[0042] After designing specific primers for the firefly luciferase (Firefly-Luciferase) gene, the Luciferase cDNA was amplified by PCR using the plasmid carrying the Luciferase gene as a template (Beiyuntian Company). The amplified Luciferase gene was successfully inserted into the PCR-Blunt vector, transformed into competent Escherichia coli cells, and the plasmid was extracted. After double digestion with Xba I and Sal I, the Luciferase gene fragment was purified and recovered by slicing, and the fragment was combined with a lentiviral eukaryotic expression vector. PRRL-CMV empty plasmid (Backbone) connection, plasmid extraction after transformation of competent E. coli cells, double enzyme digestion identification and sequencing results all proved that the Luciferase-PRRL-CMV expression vector was successfully constructed, and the sequence was confirmed to be completely correct (about 1700bp)

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Abstract

The invention relates to an A549 nude mouse model of stably expressed luciferase and building and application thereof. The nude mouse model leads firefly luciferase genes to human lung adenocarcinoma A549 cell strains through the gene recombination technology and lentivirus infection, tumor clone cell strains of the stably expressed luciferase are obtained through subcloning screening, and recombinational A549 cells are inoculated in a mouse to obtain the A549 nude mouse model. Compared with a traditional tumor model drug effect detection method, the A549 nude mouse model enables observation to be visual and convenient, can be used for somatoscopy without damaging animals, is reliable in result, and has good application prospect.

Description

technical field [0001] The invention belongs to the field of nude mouse model and its establishment and application, in particular to an A549 nude mouse model stably expressing luciferase and its establishment and application. Background technique [0002] Lung cancer is the number one killer threatening the health of our people. According to the "2010 China Health Statistical Yearbook" issued by the Ministry of Health, in 2009, malignant tumors ranked first among all causes of death in my country. Statistics show that among the top ten malignant tumors with the highest mortality rate, lung cancer ranks first. Development carries a serious economic burden. In recent years, the treatment and basic research of lung cancer have made great progress, but the current basic and preclinical research of lung cancer drugs still lacks a reliable in vivo and in vitro research platform that can more conveniently evaluate the efficacy of drugs, which seriously restricts lung cancer. basi...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/867C12N5/10A01K67/027A61K49/00
Inventor 房健民周爽杨洋李栋
Owner SUZHOU RES INST OF TONGJI UNIV
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