Document Type : Research Paper
Authors
Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran.
Abstract
Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology and to shorten the time for gene functional analysis. Transient expression is a fast, flexible, simple and easy method that could be used in fully differentiated plant tissues. 2A11 promoter is a tomato fruit specific promoter whose expression occurs in fruit significantly. Cloning of fruit-specific promoter (2A11) is the main purpose of this study. 2A11 promoters were amplified from genomic DNA of tomato by PCR using specific primers. The promoter fragments were cloned into cloning vector PTZ57R/T. Recombinant plasmids were transferred into E. coli XLI-blue strain. Fragments of the interest were digested using restriction enzymes BamH1 and HindIII and were then purified and substituted in CaMV35S promoter in binary pBI121. Binary pBI121 was selected for cloning of 2A11 promoters as it is an ideal vector for agroinfiltration due to the presence of the CaMV35S promoter and the GUS reporter gene within its T-DNA region. Recombinant plasmids were transformed into Agrobacterium tumefaciens LBA4404 strain with freeze and thawing method. The expression of GUS gene was analyzed in tomato with agroinfiltration method. The results showed that, 2A11 promoter was found as efficient as CaMV35S promoter in expression of GUS gene specifically in tomato fruit. Then, we can use this promoter for tissue specific expression of recombinant proteins in tomato fruits.
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