ISSN:
1573-6784
Keywords:
assembly PCR
;
codon usage
;
malaria
;
mycobacterium
Source:
Springer Online Journal Archives 1860-2000
Topics:
Process Engineering, Biotechnology, Nutrition Technology
Notes:
Abstract The cloning of an A:T-rich DNA fragment, the C-terminus of Plasmodium falciparum merozoite surface protein-1, into the G:C-rich Mycobacterium smegmatis resulted in delayed growth (6 days), a 10-fold decrease in the expected transformation efficiency and low level expression of the recombinant protein by the host cells. The technique of assembly PCR was used to assemble a series of oligonucleotides to generate a synthetic homologue of the plasmodium DNA fragment using mycobacterium codon bias. Cloning of the synthetic fragment into M. smegmatis resulted in normal growth (4 days), the expected level of transformation efficiency (104 c.f.u. μg−1 DNA) and a substantial increase in the expression of the recombinant protein.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1008945112490
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