Two methods for rapid assembly and oligomerization of synthetic genes: construction of human calcitonin-encoding sequences.
Two fast, simple and reliable methods for construction of synthetic genes are described. The first represents a one-vial method for assembly and cloning of monomeric genes, and the second allows a simultaneous assembly, oligomerization and cloning of synthetic genes. Both methods consist of ligation of the synthetic oligodeoxyribonucleotides (oligos) building up the gene directly into the cloning vector, followed by screening of the recombinant clones for the presence of specific gene constructs. The average length of the oligomeric genes thus constructed is directly proportional to the molar ratio between the vector DNA and the synthetic oligos used. Both methods are illustrated by the construction of a series of human calcitonin-encoding genes.
Ivanov
I
I
Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia.
Christov
C
C
Alexciev
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002116:Calcitonin / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D005813:Genes, Synthetic; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics
D009838:Oligodeoxyribonucleotides; D002116:Calcitonin