Construction of the High Throughput Technology for Screening Osmotic Stress Relevant Genes / 中国生物工程杂志

ABSTRACT

With the development of functional genomics, high throughput analysis of genes’ function has been the mainstream of research, and exogenous gene's over expression via Agrobacterium-mediated transformation is the most commonly used method in gene functional analysis.The versatile plant expression vector cassette named pBHT-5 was constructed by the method of site-specific mutagenesis based on pBI121. First of all, the restriction enzyme SfiI recognition site in trfA gene (X00713) which was relevant to plasmid replication and stability was replaced without changing its amino acid composition. And then the SfiIA,SfiIB sites were added between promoter CaMV35s and terminator NOS. The versatile plant expression vector cassette can be directly used to construct plant expression vector containing the full-length genes cloned by Clontech SMARTTM technology, which will raise the efficiency of vector construction. The result will provide basis of new genes’ high throughput screening and functional analysis, then get the new genes functioning in plant osmotic stress resistance.