Method for constructing laryngeal cancer cell line for stably expressing green fluorescent protein

A green fluorescent protein, stable expression technology, applied in the field of cell research, can solve problems such as targeted drug screening and molecular mechanism research that limit laryngeal cancer, and achieve the effect of meeting different flux requirements and short screening period
CN108949697AInactive Publication Date: 2018-12-07FIRST HOSPITAL OF SHANXI MEDICAL UNIV
4 Cites 9 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
FIRST HOSPITAL OF SHANXI MEDICAL UNIV
Publication Date
2018-12-07
Estimated Expiration
Not applicable · inactive patent

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The invention belongs to the technical field of cell research, and particularly relates to a method for constructing a laryngeal cancer cell line for stably expressing the green fluorescent protein. The method comprises the following steps: (1) designing and synthesizing a primer; (2) digesting the gene sequence of pLenti-puro carrier plasmids with BamHI and XhoI according to the gene sequence andpLenti-puro sequence of the green fluorescent protein, and connecting digested products to obtain a target carrier pLenti-puro-GFP; (3) verifying the carrier pLenti-puro-GFP with endotoxin extraction, digesting the pLenti-puro-GFP plasmids with BamHI and XhoI, and detecting the gene sequence by agarose gel electrophoresis, and sequencing the carrier pLenti-puro-GFP plasmids to detect the sequencesituation of the GFP-LC3; and (4) co-transfecting a 293T cell with the carrier pLenti-puro-GFP plasmids and a lentivirus package carrier, collecting virus supernatant, and transfecting a laryngeal cancer cell Hep-2 with the virus supernatant, infecting for 24 hours, then adding a culture medium containing puromycin, screening the cell, and observing under high content until a stable strain is obtained.
Need to check novelty before this filing date? Find Prior Art

Description

Technical field

[0001] The invention belongs to the technical field of cell research, and specifically relates to a method for constructing a laryngeal cancer cell line stably expressing green fluorescent protein. Background technique

[0002] Green fluorescent protein (GFP) was originally a protein found in jellyfish. When jellyfish luminescent protein binds to Ca 2+ Then it emits blue fluorescence, and the blue light further excites GFP to produce green fluorescence. The fluorescence emitted by GFP has a longer duration and does not require any other auxiliary factors. After the successful cloning of GFP cDNA and its successful expression in E. coli and mammalian cells, people realized its huge application prospects.

[0003] Laryngeal cancer is one of the common malignant tumors of head and neck cancer. It has the characteristics of high incidence and high recurrence rate. The incidence rate varies greatly from country to country. So far, the treatment of laryngeal cancer is st...

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