农学类论文分析.pdf

This is the Pre-Published Version
A fluorescence resonance energy transfer (FRET)-based high throughput drug screening
method for discovery of anti-pounds from herbal medicine

1Honglei Tian, 1Louisa Ip, 2Houwei Luo, 3Donald C. Chang, and *, 1Kathy Q. Luo
1Bioengineering Graduate Program, Department of Chemical Engineering, 3Department of
Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong
Kong, and 2Department of Natural Medical Chemistry, China Pharmaceutical University,
Nanjing, China

Running title: A HTS method for anti-pounds discovery

* Corresponding Author: Dr. Kathy Q. Luo, Bioengineering Graduate Program, Department
of Chemical Engineering, Hong Kong University of Science and Technology, Clear Water Bay,
Kowloon, Hong Kong, Tel: 852-2358-8434, Fax: 852-3106-4857, Email: ******@

1
Summary
Background and purpose: We report the development of a very efficient cell-based high
throughput screening (HTS) method, which utilizes a novel bio-sensor that selectively detects
apoptosis based on the fluorescence resonance energy transfer (FRET) technique.
Experimental approach: We generated a stable HeLa cell line expressing a FRET-based
bio-sensor protein. When cells undergo apoptosis, they activate a protease called “caspase-3”.
Activation of this enzyme will cleave our sensor protein and cause its fluorescence emission to
shift from a wavelength of 535 nm (green) to 486 nm (blue). A lowering of the green/blue
emission ratio thus gives a direct indication of apoptosis. The sensor cells can be grown in
96-well plates. After adding different pounds to each well, the fluorescent profile
can be measured at various times using a fluorescent plate reader. Compounds that can trigger
apoptosis are potential candidates of anti-cancer drugs.
Key results: We demonstrated that this novel cell-based HTS method is highly effective in
identifying anti-pounds. First, this s