Cloning and Characterization of Salt Stress Related Gene(s)

Abstract

Selection for high-yielding, salt tolerant varieties has proven to be an

elusive target for plant breeders and the identification of reliable genetic markers for salt tolerance has been even more elusive for plant molecular biologists. The plant is an integrated system that is adapted to specific environment on which salinity has become an intrusion. A comprehensive study to develop a salt tolerant variety should be started at the level of gene isolation and identification from halophytic plants. In this study, we isolated a full-length coding sequence of myo­ inositol !-phosphate synthase gene from Arthrocnemon glaucum plant. To accomplish this goal the following steps were performed: 1- DNA sequences for myo-inositol !-phosphate synthase gene isolated

.,, from different plants was retrieved from the GenBank.

2- The sequences were formatted and subjected to alignment usmg CLUSTL W (version 1.6) multiple sequence alignment program in the EMBL databases. 3- Conserved regions along the DNA sequences were chosen for degenerate primers to use it in the RT-PCR reaction. 4- Total RNA was isolated from Arthrocnemon glaucum plant subjected to salt stress, and then mRNA was purified from the total RNA after confirming its integrity. 5- Degenerate primers were used in the RT-PCR reaction to amplify a DNA

fragment of897 bp.

6- The amplified RT-PCR fragment was cloned and sequenced.

7- The DNA sequence was subjected to computer analysis to identify the cloned fragment.