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Bacterial libraries for improving proteins

In a bacterial library, many different DNA sequences are introduced into the cells of a bacterial culture. Libraries are powerful tools for improving protein function.

What is a bacterial library?

A bacterial library is a large number of different DNA sequences (hundreds, millions or even billions), each of which is cloned into a vector and introduced into a bacterial cell, so there are a great many different DNA sequences represented within the individual bacteria in the library – one in each bacterium. It’s called a library because it contains so many different DNA sequences – just as a public library houses a great many books.

Get information sheet: How to add foreign DNA to bacteria

One gene, many versions

One important use for bacterial libraries is in improving the function of a protein or finding a protein with a new function. To make these libraries, scientists take as their starting point a gene that encodes a protein of interest. Every DNA sequence in the library is a slightly different version of that gene (a gene variant).

The bacterial cells in the library produce protein from the gene variants. Because the variants all differ slightly in their DNA sequence, there will be many slightly different versions of the protein too. Each will have a subtly different sequence of amino acids, and each is likely to function worse or better (usually worse!) than the original protein. In some cases, a variant protein may even have acquired a different function.

To use the library, scientists test the bacteria within it to see which of the many protein variants function best.

Get focus story: Evolved enzymes

Making gene variants

Scientists use several methods to make the gene variants that make up a library. One way is to use a version of polymerase chain reaction called error-prone PCR (or sloppy PCR). In this technique, random changes to the gene’s DNA sequence are introduced each time the gene is copied. By the time the PCR is complete, billions of versions of the gene may have been produced.

Get information sheet: Polymerase chain reaction (PCR)

Bigger is better

When making a library for improving a protein, scientists usually aim to clone as many different DNA sequences as possible. Why? Let’s say that only one in a thousand random changes in DNA sequence will improve the protein’s function (that’s a fairly typical estimate). The larger the number of unique DNA sequences, the more likely it is that one of the sequences will encode an improved protein.

Currently, the biggest libraries contain many billions of unique sequences. With numbers that big, it’s not hard to see why libraries are powerful tools for finding proteins with improved function.

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