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Related Concept Videos

Protein Digestion01:02

Protein Digestion

Protein digestion begins in the stomach, where the highly acidic environment can easily disrupt protein structure by exposing the peptide bonds of polypeptide chains. After polypeptide chains are broken into individual amino acids by a series of digestive enzymes, the amino acids are transported to the liver via the bloodstream to produce energy.

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Digestion-after-shuffling: a method for combinatorial library construction.

Yingfeng An1, Shouhui Sun, Shuxia Lv

  • 1College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China. ayf66@yahoo.com.cn

Analytical Biochemistry
|May 17, 2011
PubMed
Summary
This summary is machine-generated.

We created digestion-after-shuffling, a new method for creating diverse amino acid sequences. This technique efficiently generates chimeric genes by using restriction enzymes to shuffle homologous genes after DNA shuffling.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Protein Engineering

Background:

  • Gene shuffling is a powerful technique for protein evolution and engineering.
  • Creating diverse libraries of chimeric genes is essential for discovering proteins with novel functions.

Purpose of the Study:

  • To develop a convenient and efficient method for generating diverse libraries of chimeric genes.
  • To enable the creation of recombinant plasmids with shuffled amino acid sequences.

Main Methods:

  • Developed a novel technique termed "digestion-after-shuffling" for family shuffling of amino acid sequences.
  • Utilized DNA shuffling of homologous genes followed by double digestions with specific restriction enzymes.
  • Employed gel purification to isolate fragments of expected sizes and subsequent religation to construct recombinant plasmids.

Main Results:

  • The digestion-after-shuffling method allows for the construction of chimeric genes with at least two restriction sites from different parental sequences.
  • This technique provides a convenient way to generate diverse gene libraries for protein engineering.

Conclusions:

  • Digestion-after-shuffling is an effective strategy for generating diverse gene chimeras.
  • This method facilitates the exploration of sequence space for protein evolution and optimization.