rocess works a bit like a "find-and-replace" command in your word processor. Imagine you have typed "Stephen Q. Gould" everywhere, and suddenly discover that the middle initial should be "J". The computer can be told to look for "Stephen" and "Gould" and replace what comes between them. In the same way, recombinases find recognizable sequences of DNA to the left and right of a target and replace what comes in between with the new sequence.

Homologous recombination was known to occur in bacteria, but it hadn't been possible to use it to engineer DNA, as was the case in yeast and mouse stem cells. Stewart?s team decided to try to find a strain that could do it. "We ordered as many types of E. coli as we could, looking for defects in the way they repair their DNA," he says. "After five months of work, Youming Zhang, a postdoc in the lab, found the strain."

The group quickly identified the bacterial factors involved and turned them into a new tool called Red/ET recombination that is now being adopted by biologists all over the world. It's one of the mainstays of Gene Bridges GmbH, a company that Stewart and his colleagues founded with EMBL to develop the commercial implications of the breakthrough.

"We have been pushing it to work with larger and larger bits of DNA," Testa says, "and our latest project has been to engineer an entire artificial chromosome in bacteria. We've constructed a large, complex 'cassette' that we've now inserted into a mouse in place of its normal gene."

The gene that they chose is called mixed-lineage leukemia (Mll), and is known to become defective in childhood leukemias in humans. By inserting the artificial version into the mouse, researchers hope to understand how the defects lead to disease. "There are many things that can go wrong in this gene," Testa says, "and we wanted to construct a version of it that would allow us to test as many aspects of the problem as possible."

The artificial Mll that
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Contact: Russ Hodge
hodge@embl.de
49-6221-387-252
European Molecular Biology Laboratory
24-Mar-2003

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