s100a9 recombinant adenovirus, its construction and application

A technology of S100A9, recombinant adenovirus, applied in the application, virus/phage, recombinant DNA technology and other directions, can solve the problem of lack of objective basis for the specific mechanism, and achieve the effect of stable recombinant adenovirus, convenient purchase and simple operation

Active Publication Date: 2018-03-27
THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the previous study, the method of immunohistochemistry was used to find that the expression of S100A9 was gradually increased in normal cervical tissue, cervical intraepithelial neoplasia, and cervical squamous cell carcinoma, but the specific mechanism of S100A9 in the occurrence and development of cervical cancer still lacks objective evidence.

Method used

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  • s100a9 recombinant adenovirus, its construction and application
  • s100a9 recombinant adenovirus, its construction and application
  • s100a9 recombinant adenovirus, its construction and application

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Embodiment Construction

[0032] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention and implement it, but the examples given are not intended to limit the present invention.

[0033] 1. Construction and identification of recombinant adenovirus vector plasmid pHBAd-MCMV-RFP-S100A9

[0034] 1. Digestion

[0035] attached figure 1 Is the schematic diagram of the pHBAd-MCMV-RFP overexpression vector and the key points therein.

[0036] not I and Nsi I is the insertion site, for pHBAd-MCMV-RFP vector not I and Nsi I double enzyme digestion, the enzyme digestion system is as follows:

[0037] 20μL enzyme digestion system 37℃ for 3h

[0038] 2μL

Carrier (400ng / μL)

1μL

not I

1μL

Nsi I

2μL

10×buffer

14μL

h 2 o

[0039] 2. Gel recovery and purification

[0040] (1) After DNA electr...

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Abstract

The invention discloses a S100A9 recombinant adenovirus, its construction and application. Specifically, the present invention discloses a S100A9 recombinant adenovirus vector plasmid: the S100A9 protein-encoding gene is recombined into the pHBAd-MCMV-RFP vector to obtain the S100A9 recombinant adenovirus vector plasmid, and the sequence of the S100A9 protein-encoding gene is shown in SEQ ID NO:1 shown. S100A9 recombinant adenovirus vector plasmid and backbone plasmid were transfected into HEK293 cells to obtain S100A9 recombinant adenovirus Ad‑S100A9. The recombinant adenovirus Ad‑S100A9 was transformed into cervical squamous cell carcinoma C33A cells, and the effect of S100A9 on the proliferation, invasion and migration of cervical squamous cell carcinoma cells was studied.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a recombinant adenovirus, in particular to a S100A9 recombinant adenovirus, its construction and application. Background technique [0002] Among the malignant tumors that seriously endanger women's health, the incidence of cervical cancer ranks second only to breast cancer. According to global statistics, there are about 500,000 new cases of cervical cancer every year, of which developing countries account for more than 80%. And in recent years, the incidence of cervical cancer tends to be younger. A large number of studies have shown that persistent high-risk human papillomavirus (HPV) infection is the primary factor in the incidence of cervical cancer, but most women with HPV infection do not develop cervical cancer. It shows that the occurrence and development of cervical cancer is a multi-factor, multi-step, multi-gene process, but the specific mechanism of occurrence and develop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/861C12N15/66C12N7/01C12N5/10
Inventor 朱雪琼陈苗苗王颖沈奇
Owner THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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