Plasmid expressed by PSA promoter mediated firefly luciferase gene

A technology of luciferase gene and luciferase, applied in the field of plasmids in which PSA promoter mediates the expression of luciferase gene

Inactive Publication Date: 2015-05-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Compared with luciferase-based bioluminescent technology, the sensitivity of in vivo detection is higher, and 10 2 order of magnitude cells, while fluorescence imaging techniques can only detect about 10 6 order of magnitude cells

Method used

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  • Plasmid expressed by PSA promoter mediated firefly luciferase gene
  • Plasmid expressed by PSA promoter mediated firefly luciferase gene
  • Plasmid expressed by PSA promoter mediated firefly luciferase gene

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Experimental program
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Embodiment 1

[0012] 1. Cell lines and experimental animals The human PCa cell line LNCaP was purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences; the human embryonic kidney 293T cell line was provided by the Zhejiang Key Research Laboratory of Organ Transplantation; SCID mice were purchased from Slack Laboratory Animals LLC.

[0013] 2. Main reagents RPMI-1640 medium and fetal bovine serum were purchased from Gibco; pGL3-Basic Vector, pRL-TK Vector and Luciferase Dual Luciferase Reporter Gene Kit were purchased from Promega; pZsGreen1-1 Vector was purchased from clontech ; BCA protein quantification kit, RIPA protein lysate was purchased from Beyontian Company; pMD 18-T Vector was purchased from Takara Company; lipofectamine 2000 was purchased from Invitrogen Company; antibiotic G418 was purchased from Sigma Company; luciferase substrate D-Luciferin was purchased from from Biotium Corporation.

[0014] 3. Cell culture The human PCa cell...

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Abstract

The invention discloses a plasmid expressed by prostate-specific antigen (PSA) promoter mediated firefly luciferase gene. The plasmid is formed by connecting a PSA promoter pPSA-ZsGreen dominant skeleton with a firefly luciferase gene fragment. The plasmid is applied to the transfection of prostate cancer cells to lay a foundation for constructing a prostate PSA fluorescence reporter gene animal model. The part of the tumor in the prostate cancer animal model is positioned by use of fluorescence, and then a reliable animal model can be provided for the treatment of the prostate cancer and the development of new drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a plasmid in which a PSA promoter mediates luciferase gene expression. Background technique [0002] Prostate cancer is one of the malignant tumors of the male reproductive system. It mainly occurs among the elderly in western developed countries. In recent years, the incidence rate in my country has shown an upward trend. Prostate-specific antigen (PSA) is a specific protein secreted by prostate epithelial cells. Changes in its content in serum can reflect the occurrence, development, invasion, metastasis, and prognosis of prostate cancer. It is the main sensitive index for clinical diagnosis, treatment and prognosis of cancer. PSA mainly exists in prostate cells and tissues. Under normal circumstances, the content of serum is low. When suffering from prostate cancer, due to the massive proliferation and rupture of tumor cells, PSA is released into the blood in large quantities, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
Inventor 姜海毛祺琦林奕伟孔德波杨凯
Owner ZHEJIANG UNIV
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