Construction of a genomic DNA library with a TA vector and its application in cloning of the phytoene synthase gene from the cyanobacteriumSpirulina platensis M-135

Abstract

An efficient and simple method for constructing a genomic DNA library using a TA cloning vector is presented. It is based on the sonicative cleavage of genomic DNA and modification of fragment ends withTaq DNA polymerase, followed by ligation using a TA vector. This method was applied for cloning of the phytoene synthase genecrt B fromSpirulina platensis. This method is useful when genomic DNA cannot be efficiently digested with restriction enzymes, a problem often encountered during the construction of a genomic DNA library of cyanobacteria.