[Methods for construction of transgenic plant expression vector: a review]

Area of Science:

• Molecular Biology
• Plant Biotechnology
• Genetic Engineering

Background:

• Recombinant plasmid vector construction is essential for transgenic plant development.
• Choosing the right method impacts cost-effectiveness and plasmid diversity.
• Traditional methods include Gateway, three DNA, and one-step cloning.

Purpose of the Study:

• To summarize working experience and published reports on DNA recombination methods for plant transformation.
• To disseminate knowledge on widely used and novel vector construction techniques.
• To provide insights into efficient and diverse recombinant plasmid creation.

Main Methods:

• Oligonucleotide synthesis-based construction for small gene expression vectors.
• Pre-microRNA based construction of small RNA expression vectors.
• Recombination-fusion PCR and In-Fusion Kit for inserting DNA fragments.
• Sequence and ligation-independent cloning, Gibson assembly, and Golden Gate assembly for complex vectors.

Main Results:

• Novel methods offer efficient construction of small fragment and small RNA expression vectors.
• Recombination-fusion PCR and In-Fusion Kit facilitate insertion of multiple restriction sites and fragments.
• Advanced assembly methods enable the construction of complex vectors with numerous fragments.

Conclusions:

• A variety of DNA recombination methods are available for plant transformation vector construction.
• Novel techniques enhance efficiency, cost-effectiveness, and diversity in creating recombinant plasmids.
• Understanding these methods is key for advancing plant genetic engineering and gene function studies.