4.2. Construction of Plant Expression Vector

IFS cDNA (Genbank accession number: {"type":"entrez-nucleotide","attrs":{"text":"AF195798","term_id":"6979519","term_text":"AF195798"}}AF195798) was kindly provided by Dr. Oliver Yu and Dr. Brian Mc Gonigle, Donald Danforth Plant Science Center, USA. The IFS gene was cloned into plant expression vector pEarleyGate 102 HA and pEAQ-HT-DEST 3 by gateway cloning technology (Invitrogen, Carlsbad, CA, USA) as per manufacturer’s protocol. The IFS gene was amplified by PCR, and purified PCR products were cloned into entry vector pDONR/Zeo and moved to destination vectors pEarleyGate 102 HA and pEAQ-HT-DEST 3, as described previously [21]. The transformed colonies in the destination vector were confirmed by gene-specific PCR and restriction digestion. Subsequently, the plant expression vectors were transformed to A. tumefaciens strain AGL1 by electroporation.