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Edited on Sat Nov-15-03 06:43 AM by treepig
of work that's been underway for 25 years now. the basic concept of using synthetic oligonucleotides for gene assembly was reported in 1980:
Science. 1980 209(4463):1401-5. Chemical DNA synthesis and recombinant DNA studies.
Chemically synthesized DNA has been used in many recombinant DNA studies. These uses have included the total synthesis and cloning of functional genes, the cloning and expression of natural genes, and editing of changing genes by directed mutation.
this technology was applied to a specific gene two years later:
J Biol Chem. 1982 257(16):9226-9. An alternate method for synthesis of double-stranded DNA segments.
Recent progress in the chemical synthesis of DNA has now made it possible to rapidly synthesize single-stranded DNAs over 40 bases in length. We have taken advantage of these longer DNAs in assembling and cloning a 132-base pair gene segment coding for amino acids 126 through the stop codon of human leukocyte interferon alpha 2. . . . We describe in detail this methodology for the biochemical assembly of long gene segments from synthetic oligodeoxyribonucleotides.
by incorporation of PCR into the protocols, much longer gene sequences were being made eight years ago:
Gene. 1995 164(1):49-53. Single-step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides.
Here, we describe assembly PCR as a method for the synthesis of long DNA sequences from large numbers of oligodeoxyribonucleotides (oligos). The method, which is derived from DNA shuffling , does not rely on DNA ligase but instead relies on DNA polymerase to build increasingly longer DNA fragments during the assembly process. A 1.1-kb fragment containing the TEM-1 beta-lactamase-encoding gene (bla) was assembled in a single reaction from a total of 56 oligos, each 40 nucleotides (nt) in length. . . We tested the range of assembly PCR by synthesizing, in a single reaction vessel containing 134 oligos, a high-molecular-mass multimeric form of a 2.7-kb plasmid containing the bla gene, the alpha-fragment of the lacZ gene and the pUC origin of replication. Digestion with a unique restriction enzyme, followed by ligation and transformation in Escherichia coli, yielded the correct plasmid. Assembly PCR is well suited for several in vitro mutagenesis strategies.
and now, from the boston globe article in post #10 - the statement "A team of scientists announced yesterday it has found a fast and accurate way to build genes from scratch, a technique that could give scientists the practical tools to create life in a lab" is just flat out a lie.
in reality, venter and associates did not find a fast and accurate way to build genes from scratch, in fact eight years ago a plasmid almost as large had been made by similar methodology (see the "gene" paper listed above) - they merely applied well-established techniques to a new DNA sequence (the bacteriophage). venter is a shameless self-promoter, but still, the globe should know better than publish such a misleading article.
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