Construction of a fulllength cDNA library of Emp

Authors: Guohua Sun Xiangquan Liu Lihua Ren Jianmin Yang Xiumei Wei Jialong Yang

Publish Date: 2013/11/08

Volume: 13, Issue: 1, Pages: 169-173

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Abstract

The basic genetic characteristics important functional genes and entire transcriptome of Solen grandis Dunker were investigated by constructing a fulllength cDNA library with the ‘switching mechanism at the 5′end of the RNA transcript’ SMART technique Total RNA was isolated from the immunerelevant tissues gills and hemocytes using the Trizol reagent and cDNA fragments were digested with Sfi I before being ligated to the pBluescript II SK vector The cDNA library had a titer of 1048 cfu μL−1 and a storage capacity of 105×106 cfu Approximately 98 of the clones in the library were recombinants and the fragment lengths of insert cDNA ranged from 08 kb to 30 kb A total of 2038 expressed sequence tags were successfully sequenced and clustered into 965 unigenes BLASTN analysis showed that 240 sequences were highly similar to the known genes Evalue 1e −5 percent identity 80 accounting for 25 of the total unigenes According to the Gene Ontology these unigenes were related to several biological processes including cell structure signal transport protein synthesis transcription energy metabolism and immunity Fifteen of the identified sequences were related to defense and immunity The fulllength cDNA sequence of HSC70 was obtained The cDNA library of S grandis provided a useful resource for future researches of functional genomics related to stress tolerance immunity and other physiological activities

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