DNA shuffling, in contrast, is PCR without synthetic primers. In this process, a family of related genes--say, the ones that codes for the surface proteins of three different HIV isolates--are digested with restriction enzymes. The gene fragments then are heated up to separate them into single-stranded templates. Some of these fragments will bind to other fragments that share complementary DNA regions, which in some cases will be from other family members. Regions of DNA that are non-complementary hang over the ends of the templates (see illustration). The PCR reaction then treats the complementary regions as primers and builds the new double-helical DNA. But PCR also adds bases to the overhanging piece of the primer, forming a double helix there, too. This ultimately creates a mixed structure or chimera. In the final step, PCR reassembles these chimeras into full-length, shuffled genes.
References
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Last updated on Monday September 29, 2008 at 10:12:10 PDT (GMT -0700)
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DNA shuffling, in contrast, is PCR without synthetic primers. In this process, a family of related genes--say, the ones that codes for the surface proteins of three different HIV isolates--are digested with restriction enzymes. The gene fragments then are heated up to separate them into single-stranded templates. Some of these fragments will bind to other fragments that share complementary DNA regions, which in some cases will be from other family members. Regions of DNA that are non-complementary hang over the ends of the templates (see illustration). The PCR reaction then treats the complementary regions as primers and builds the new double-helical DNA. But PCR also adds bases to the overhanging piece of the primer, forming a double helix there, too. This ultimately creates a mixed structure or chimera. In the final step, PCR reassembles these chimeras into full-length, shuffled genes.
References
This article is licensed under the GNU Free Documentation License.
Last updated on Monday September 29, 2008 at 10:12:10 PDT (GMT -0700)
View this article at Wikipedia.org - Edit this article at Wikipedia.org - Donate to the Wikimedia Foundation
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