Gene construction advice needed

Anders Gorm Pedersen gorm at cbs.dtu.dk
Mon Jul 10 02:26:24 EST 2000


"Joseph C. Bagshaw" wrote:
> 
> I'm looking for advice on a gene construction project.  I'm trying to
> assemble a 219 bp gene from oligonucleotides.  I designed the oligos so
> that each overlaps the next by ten complementary bases.  The plan then is
> to fill the gaps, then ligate.  

Here are some literature pointers that might be of help:

Single-step assembly of a gene and entire plasmid from large numbers of
oligodeoxyribonucleotides.
Stemmer WP, Crameri A, Ha KD, Brennan TM, Heyneker HL
Gene 1995 Oct 16 164:1 49-53

Gene synthesis by a LCR-based approach: high-level production of
leptin-L54 using synthetic gene in Escherichia coli.
Au LC, Yang FY, Yang WJ, Lo SH, Kao CF
Biochem Biophys Res Commun 1998 Jul 9 248:1 200-3

And some older ones:

Dual asymmetric PCR: one-step construction of synthetic genes.
Sandhu GS, Aleff RA, Kline BC
Biotechniques 1992 Jan 12:1 14-6

Two methods for rapid assembly and oligomerization of synthetic genes:
construction of
human calcitonin-encoding sequences.
Ivanov I, Christov C, Alexciev K
Gene 1990 Nov 15 95:2 295-9

Construction of synthetic genes using PCR after automated DNA synthesis
of their entire top and bottom strands.
Ciccarelli RB, Gunyuzlu P, Huang J, Scott C, Oakes FT
Nucleic Acids Res 1991 Nov 11 19:21 6007-13


Best regards,
Anders Gorm
---------------------------------------------------------------
 Anders Gorm Pedersen, cand.scient., Ph.D.  (gorm at cbs.dtu.dk)

 Center for Biological Sequence Analysis
 Technical University of Denmark
 Bldg 208, DK-2800 Lyngby, Denmark

 phone: (+45) 45 25 24 84
 fax:   (+45) 45 93 15 85

 Web: http://www.cbs.dtu.dk/gorm/
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