EFFICIENT LIBRARY CONSTRUCTION WITH A TA VECTOR AND ITS APPLICATION TO CLONING OF THE PHYTOENE SYNTHASE GENE FROM THE CYANOBACTERIUM SPIRULINA-PLATENSIS

An efficient and simple method for constructing a genomic DNA library is presented by use of a TA cloning vector. It is based on sonicative cleavage of genomic DNA and modification of the fragment ends with Tag DNA polymerase, followed by ligation with a TA vector. This method wa s successfully applied to cloning of the phytoene synthase gene crtB f rom Spirulina platensis. The method is useful when the genomic DNA is not well digested with restriction enzymes owing to methylation or oth er reasons.