To date, genes have only been used for expression in bacteria, so it is not known whether this simple strategy could also work for transgenesis in higher plants. The software program in CAPITALS is from the DOS Compugene suite of DNALYSIS software, which can be obtained from the author (W.M.B) running under Windows. This experiment was derived from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu Lijia.
Operation method
Codon design experiment Move 1. In order to emulate the codon frequency of a high level expressed plant gene, the codons of the small subunit of tomato Rubisco (ribulose bisphosphate carboxylase/oxygenase) were calculated using the CODONS program. Unfortunately, this single gene was probably too small and too extreme a sample, as it did not have a sample of every codon. A codon list for another gene that is expressed at high levels in plants [the gene for the tobacco mosaic virus capsid protein (TMV)] and a codon list for a gene from a bacterial source that is known to have no problems with gene expression in plants [that is, the NPT-II (kanamycin resistance) gene from Tn5] were then added. the CODONS program accepted these genes in order and then automatically created a master codon table. For more product details, please visit Aladdin Scientific website.
2. Next, a list of amino acids for the target proteins Ds-Red and CryV was generated in single-letter code using the TRAFIG program. This output file is edited into a standard fasta form and is then used as the input file for the REVTRANS program, which requires not only the amino acid sequence, but also a codon table to mimic it. REVTKANS can easily read in the output format of the master list created by CODONS. The sequence generation program is somewhat slower (no matter how fast the computer is running) because it produces only 18 codons per second. This is because it takes 1/18s from the new year to the latest to seed a pseudo-random number generation program. the REVTRANS program can be run continuously, each time yielding a different (partially homologous) DNA sequence, but each sequence encodes the input protein and has almost identical codons in their codon tables as required by the user. The codon usage frequency is modeled after the user's requirements.