Catch & Release-rapid cost-effective protein purification from plants using a DIY GFP-Trap-protease approach

Area of Science:

• Plant molecular biology
• Protein biochemistry
• Biotechnology

Background:

• Protein purification is essential for studying protein structure and function.
• Plant protein purification faces challenges like complex tissues, secondary metabolites, and low protein abundance.
• Conventional methods are often expensive, multistep, and labor-intensive.

Purpose of the Study:

• To present a cost-effective, fast, and reliable one-step protein purification workflow for plant tissues.
• To introduce the 'Catch & Release' system, including a vector set, homemade GFP-Trap, and proteases.
• To streamline cloning and transgenic plant selection using fluorescence-accumulating seed technology.

Main Methods:

• Developed a modular vector set with interchangeable plant promoters, cloning dropout markers, protease cleavage sites, and epitope tags.
• Utilized fluorescence-accumulating seed technology for efficient transgenic plant selection.
• Employed homemade GFP-Trap and proteases for protein isolation from plant tissues.

Main Results:

• Demonstrated streamlined cloning and transgenic plant selection via fluorescence-accumulating seeds.
• Confirmed in vivo functionality of vector modules through complementation and transient expression.
• Achieved efficient and rapid isolation of enzymatically active, soluble, and high molecular weight membrane proteins.
• Reduced purification costs by up to 60-fold compared to commercial options.

Conclusions:

• The 'Catch & Release' system offers a flexible and efficient solution for plant protein purification.
• This method significantly reduces costs and labor associated with protein isolation from plants.
• The system facilitates diverse molecular biology applications and protein research in plants.