EcoRI

Definition

EcoRI is a type of restriction enzyme, a protein that can recognize and cleave specific DNA sequences, allowing for the manipulation and analysis of genetic material. It is a crucial tool in the field of DNA sequencing and genetic engineering.

5 Must Know Facts For Your Next Test

1. EcoRI is isolated from the bacterium Escherichia coli and recognizes the palindromic DNA sequence 5'-GAATTC-3'.
2. When EcoRI binds to this recognition sequence, it creates a staggered cut, leaving 'sticky ends' that can be used in DNA cloning and recombination.
3. EcoRI is one of the most commonly used restriction enzymes in molecular biology and genetic engineering due to its reliable and predictable cleavage pattern.
4. The use of EcoRI allows for the precise excision and isolation of DNA fragments, which is essential for techniques like DNA sequencing, gene cloning, and genetic mapping.
5. EcoRI is a type II restriction enzyme, meaning it recognizes and cleaves a specific DNA sequence without the need for any additional cofactors or energy sources.

Review Questions

• Explain the role of EcoRI in the process of DNA sequencing.
  • EcoRI plays a crucial role in DNA sequencing by allowing for the precise excision and isolation of DNA fragments. The enzyme's ability to recognize and cleave the specific palindromic sequence 5'-GAATTC-3' enables researchers to generate manageable, sequenceable DNA fragments from larger genomic DNA. These fragments can then be individually sequenced and assembled to determine the complete DNA sequence of a gene or entire genome, a fundamental step in the DNA sequencing process.
• Describe how the recognition and cleavage pattern of EcoRI contributes to its utility in genetic engineering and recombinant DNA technology.
  • The recognition sequence of EcoRI, 5'-GAATTC-3', is a palindromic sequence, meaning it reads the same forward and backward. This allows EcoRI to create a staggered cut, leaving 'sticky ends' on the resulting DNA fragments. These sticky ends can be used to ligate, or join, DNA fragments from different sources, enabling the creation of recombinant DNA molecules. This process is essential for a wide range of genetic engineering applications, such as gene cloning, genetic modification, and the construction of plasmids and other vectors for gene expression and manipulation.
• Evaluate the significance of EcoRI's classification as a type II restriction enzyme and how this contributes to its widespread use in molecular biology.
  • EcoRI is classified as a type II restriction enzyme, which means it recognizes and cleaves a specific DNA sequence without the need for any additional cofactors or energy sources. This makes EcoRI a highly versatile and reliable tool in molecular biology and genetic engineering. Unlike some restriction enzymes that require specific conditions or co-factors, EcoRI can be used in a wide range of experimental setups and protocols, simplifying the process of DNA manipulation and analysis. This, combined with its predictable and consistent cleavage pattern, has made EcoRI one of the most commonly used restriction enzymes in the field, facilitating numerous advancements in areas such as gene cloning, genome mapping, and DNA sequencing.

Related terms

Restriction Enzyme: Enzymes that can recognize and cut DNA at specific sequences, used in genetic engineering and molecular biology.

DNA Cleavage: The process of breaking apart DNA molecules at specific sites using restriction enzymes.

Palindromic Sequence: A DNA sequence that reads the same forward and backward, which is the recognition site for many restriction enzymes like EcoRI.