Construction of cDNA Library from Leaves of Bruguiera gymnorrhiza Seedlings and Its Expressed Sequence Tags Analysis

Abstract:

The seedlings of Bruguiera gymnorrhiza were cultured under 25‰ salinity for 4 months, then the total RNA were extracted from leaves. Following the protocol of SMARTTM cDNA Library Construction kit, the higher quality cDNA library was constructed. The primary titer of the cDNA library was about 1×106 cfu mL-1, its recombinant efficiency was calculated as 94.4%, and the size of the insert DNA fragments was ranged from 1 kb to 2 kb. Ninety-six randomly selected recombinant clones from the cDNA library were sequenced to produce 94 expressed sequence tags (ESTs), among which 81 unigenes (including 5 contigs and 76 singletons) were generated after quality control and assembly of sequences, they were all homologous to the sequences in NCBI Nr database by blastx analysis (E<10-5). These unigenes involved in the processes of respiration, photosynthesis, glucose metabolism, amino acid metabolism, fatty acid metabolism, biosynthesis of unsaturated fatty acids, etc, and had relation to the damage repair, endocytosis, and PPAR signaling pathway.

Key words:Bruguiera gymnorrhiza; Leaf; cDNA library; Expressed sequence tags; Functional analysis