Construction of a cDNA Library from the Ephemeral Plant Olimarabidopsis pumila and Preliminary Analysis of Expressed Sequence Tags

Abstract

Olimarabidopsis pumila is a close relative of the model plant Arabidopsis thaliana but, unlike A. thaliana, it is a salt-tolerant ephemeral plant that is widely distributed in semi-arid and semi-salinized regions of the Xinjiang region of China, thus providing an ideal candidate plant system for salt tolerance gene mining. A good-quality cDNA library was constructed using cap antibody to enrich full-length cDNA with the gateway technology allowing library construction without traditional methods of cloning by use of restriction enzymes. A preliminary analysis of expressed sequence tags (ESTs) was carried out. The titers of the primary and the normalized cDNA library were 1.6 · 10 6 cfu/mL and 6.7 · 10 6 cfu/mL, respectively. A total of 1093 clones were randomly selected from the normalized library for EST sequencing. By sequence analysis, 894 high-quality ESTs were generated and assembled into 736 unique sequences consisting of 72 contigs and 664 singletons. The resulting unigenes were categorized according to the gene ontology (GO) hierarchy. The potential roles of gene products associated with stress-related ESTs are discussed. The 736 unigenes were similar to A. thaliana, A. lyrata, or Thellungiella salsuginea. This research provides an overview of the mRNA expression profi le and fi rst-hand information of gene sequence expressed in young leaves of O. pumila.

Generation and analysis of expressed sequence tags from roots cDNA library of cotton (Gossypium arboreum)

“…The titer of the original and amplified library of the cotton roots were 1.22×10 6 PFU mL -1 and 10 8 PFU mL -1 respectively. Earlier studies reported that titer of cDNA library of root 1.6×10 6 PFU mL -1 (Zhao et al, 2013) which support our results. The data generated by EST's of randomly selected transcripts isolated under specific stress factors provide a chance to explore gene function and their category besides their different physiological progressions in plants against different stresses (Wong et al, 2005).Genomic approaches may help in understanding the physiological events at cellular and molecular level by dissecting the genetic resources of plants (Chaves et al, 2003).…”