When it's about molecular biology, precision and accuracy are crucial for studying the structure and function of cellular molecules such as proteins, nucleic acids, and lipids. By joining two DNA fragments to establish a new chemical bond, enzymes are responsible for ensuring this precision. There are various types of ligase enzymes, including DNA ligase, T4 DNA ligase, MDM2/mdmx ligase, and RING-finger E3 ligase. In this blog, we will explore the role of T4 DNA ligase in DNA manipulation. Among these, T4 DNA ligase is the most extensively used ligase in molecular biology. In this blog, we will explore the role of T4 DNA ligase in DNA manipulation.
What Exactly is T4 DNA Ligase?
T4 DNA ligase is an enzyme involved in catalyzing the formation of a phosphodiester bond. The enzymes can efficiently joint blunt and cohesive ends of DNA ligation and repair single-stranded nicks in duplex DNA and some DNA/RNA hybrids, facilitating the formation of a continuous DNA strand. T4 DNA ligase has obtained popularity in various routine applications ranging from molecular cloning, library construction, and high-throughput DNA sequencing to the detection of small RNAs. Due to its versatility of ligating juxtaposed 5' phosphate and 3' hydroxyl termini in duplex DNA or RNA, it has been optimized for a range of conditions for fast and efficient ligation.
Working Mechanism of T4 DNA Ligase
The process of DNA ligation includes three steps:
Key Points about T4 DNA Ligase
Applications of T4 DNA Ligase
These enzymes can be used for a variety of applications in molecular biology, including:
DNA Cloning: T4 DNA ligase is essential in the cloning process where both blunt and cohesive-ended double-stranded DNA molecules are ligated, usually between a plasmid vector and insert. They can be used in molecular cloning of PCR products or restriction fragments.
Gene Editing: In gene editing, T4 DNA ligase helps in enhancing in vitro activity in lab workflows. Also, this ligase can be used to exploit new genetic engineering tools and is expected to promote genome streamlining. T4 DNA ligase can be used in combination with CRISPR in bacterial genome engineering.
Self-Circularization: T4 DNA ligases are used in the self-circularization of linear DNA. For instance, Cytiva’s Ready-To-Go T4 DNA Ligase includes self-circularization even prior to transformation, ligation of the insert to vector, and ligation of synthetic linkers to blunt-ended DNA.
Genetic Engineering: In genetic engineering, T4 DNA ligase is used to insert DNA fragments into the genomes of organisms, enabling the production of genetically modified plants, animals, and microorganisms.
What are Limitations?
Top companies in T4 DNA Ligase Market:
Recent Product Launches
In June 2024, New England Biolabs and Inorevia collaborated to maximize data quality by preparing sequence-ready libraries from challenging samples using NEBNext library preparation reagents on Inorevia's Magelia automation platform.
In February 2023, New England Biolabs released the NEBNext UltraExpress DNA and RNA Library Prep Kits for next-generation sequencing on the Illumina platform. These new kits enable RNA library preparation in a single day and DNA library preparation in under two hours with high-quality libraries and a faster workflow. New England Biolabs is a US-based life sciences company that specializes in producing and delivering recombinant and native enzyme reagents for life science research.
In August 2015, Merck KGaA, a German-based multinational science and technology company, expanded its portfolio of enzymes for molecular biology applications by launching the “Rapid DNA Ligation Kit” featuring ready-to-use reagents to ensure accuracy and reproducibility.
Final Thoughts
The field of ligase enzymes is expected to experience significant growth in upcoming years as biotechnology companies and academic institutions continue to collaborate to revolutionize T4 DNA Ligase technology. Such initiatives reflect the potential development of new enzyme variants that are pivotal in emerging applications within genomics and biotechnology. However, the T4 DNA enzyme will undoubtedly remain a key player in reshaping molecular biology.
Despite these advancements, T4 DNA Ligase is likely to remain a key player in molecular biology. Developments in the biotechnology and pharmaceutical sectors, such as advancements in genetic and protein engineering, have expanded the applications of enzymes and are anticipated to offer lucrative growth opportunities throughout the projected period.
Attainment of client satisfaction being our motto, we at Polaris are happy to offer you multitude of reasons for why you should select us for your businesses.
Our commitment to our clients lies not only from the inception of our pre-on boarding discussions but also beyond the completion and delivery of our services.
We categorically offer custom-made/ tailored services for our syndicated research reports to provide you insights to meet the strategic requirements of your organization.
On lines of the industry best-practices, we offer our clients the finest in research and consulting services with the most reliable and accurate research findings.