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Adenovirus chip

An adenovirus and chip technology, applied in the field of biomedicine, can solve the problems of large consumption of reagents, increased mutual pollution, and high product prices, and achieve the effects of less mutual pollution, reliable data, and reduced differences

Active Publication Date: 2014-03-19
SHANDONG VIGENE BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Low transfection efficiency ultimately affects protein expression and cell function research
[0009] 2. Existing chip technology cannot be used to study non-dividing cells
[0010] 3. When the density of the sample point on the plasmid gene chip increases, the mutual contamination will increase
[0011] 4. The existing high-throughput analysis of full-length genes has low efficiency and consumes a lot of reagents
[0012] 5. The price of prior art products is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Preparation of adenovirus chip

[0045] (1). Vector preparation

[0046] General microscope slides can be used as carriers. Slides were first immersed in 25 μg / mL nitrocellulose in methanol, then allowed to evaporate the methanol under aseptic conditions, and dried.

[0047] (2). Adenovirus preparation

[0048] It mainly includes the preparation of adenovirus plasmid vector and the packaging production of adenovirus.

[0049] 1) Each human gene clone is first amplified with ORF primers, cloned into a shuttle vector by a high-throughput method, and then recombined into an adenovirus backbone vector by a high-throughput method.

[0050] 2) Packaging of adenovirus. After linearization, the adenovirus carrying the human gene was added to the wells of a 6-well plate together with the transfection reagent, and cultured for 13 days. Afterwards, the cells were scraped off, centrifuged at a low speed of 2000 rpm, added 1 ml of PBS buffer, and frozen and thawe...

Embodiment 2

[0055] Example 2. Converting Adenovirus Chips to Protein Chips

[0056] Put the adenovirus chip into the petri dish, add 1×10 6 Cells (such as HEK293 cells, CHO cells), 8-10 hours, 37 degrees, 5% carbon dioxide culture. Then add fresh culture medium. According to different experimental requirements, the cells can be treated differently.

[0057] The manipulation of cells is a common skill in the art.

Embodiment 3

[0058] Example 3 . Addition and detection of fluorescent markers on adenovirus chip

[0059] (1) When making shuttle vectors and adenovirus clones, the foreign gene and green fluorescent protein (GFP) are healed together. When cells are added to the chip and cells are transfected, fluorescence can be directly observed under a fluorescence microscope. Alternatively, GFP can also be expressed in the same vector with different promoters or by using IRES. Both fluorescence and color can be obtained with high-throughput scanning microscopy techniques.

[0060] (2) Use different labels or pigments to mark small molecular compounds, proteins or other substances for detection. For example, to measure the binding of a cell surface receptor (Receptor) and a ligand (Ligand), the ligand can be labeled with different fluorescent substances or pigments. When adding cells to the adenovirus chip, the labeled ligand is also added at the same time. When a ligand binds to a receptor in th...

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Abstract

The invention belongs to the field of biomedicines, relates to an adenovirus chip and an application thereof and particularly relates to an adenovirus chip containing a human full-length genome and an application of the adenovirus chip. The adenovirus chip provided by the invention is prepared by printing adenovirus particles containing human full-length genes on a microscope glass chip. High-concentration adenovirus particles are produced, and high-concentration adenovirus is printed on a nitrated fiber glass sheet or other substrates by using a chip microarrayer. The adenovirus chip provided by the invention can also be converted into a protein chip. The adenovirus chip provided by the invention can be applied to multiple different biomedical researches including living cell analysis (cell growth and differentiation, cell apoptosis, tissue specificity genetic expression and analysis of cells to toxicity and sensitivity of micromolecules) and can also be used for immunity analysis, enzymatic activity analysis and promoter / reporter gene analysis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to an adenovirus chip and its application, in particular to a human whole genome adenovirus chip and its application. Background technique [0002] After the entire human genome has been sequenced, the most important next step is how to discover the function of each gene. Only by knowing the function of each gene can we effectively treat various human diseases. Every human gene performs a certain function in the body, and the occurrence of diseases, especially cancer, is related to genes. In order to study the function of each gene, a specific cDNA gene clone must first be obtained. Gene cloning circulates like "banknotes" in the biotechnology and biopharmaceutical industries. Because the emergence and development of many diseases are the result of the joint action of many genes, future functional genomics, proteomics, and new drug development will rely on the simultaneous use of many kin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12Q1/02G01N33/68C12N15/861
Inventor 孙在仁
Owner SHANDONG VIGENE BIOSCI
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