中国水产科学2011，Vol.18Issue(1)：222-229,8.DOI:10.3724/SP.J.1118.2011.00222
虾夷马粪海胆性腺cDNA文库构建及部分EST序列分析
Construction of cDNA library and partial ESTs analysis of Strongylocentrotus intermedius
摘要
Abstract
Sea urchin(Strongylocentrotus intermedius) is a species of commercial and ecological significance, and as a economic species, sea urchin has been demanded steadily in Europe and Southeastern area for a long time.Recent studies focused on the development of its genomic resources, which is a key step towards further investigation, identification of gene and gene networks involved in its economic characters. Efforts have been focused on genes that can affect expression of important economic traits, such as growth and nutritional traits related genes. In this study, a cDNA library of sea urchin has been constructed from gonad using Gubler-Hoffman cDNA library technique. Total RNA was extracted from the grounded frozen powder of gonad tissue by using acid-guanidinephenol-chloroform (AGPC). Poly(A)+ RNA was isolated and purified by oligo(dT)18 anchor primer containing Not Ⅰ site. Both ends of the newly generated and polished double-stranded (ds) cDNA were attached by EcoR Ⅰ adaptors and the dscDNA was then restricted by Not Ⅰ. The short cDNA (＜400 bp) was removed by Spin Column. The library consisted of 2.10× 106 PFU/mL colony forming units with average insert size of 1.4 kb, ranging from 0.5 kb to 4.2 kb and was quantified to construct a cDNA library. Eight hundred and fourteen ESTs were compared with sequences in GenBank database of NCBI using Blast search, among which 65 ESTs and cDNA clones had been identified. These identified ESTs were submitted to GenBank with accession number of GO448010-GO448016 and GR410172-GR410229. The cDNA library provides rich source for better understanding of functional genes in sea urchin, and thus will play an important role in further development of sea urchin research.关键词
虾夷马粪海胆/cDNA/表达序列标签分类
生物科学引用本文复制引用
丁君,孙巍,常亚青..虾夷马粪海胆性腺cDNA文库构建及部分EST序列分析[J].中国水产科学,2011,18(1):222-229,8.基金项目
国家863计划项目(2006AA10A411) （2006AA10A411）
国家自然科学基金项目(30972269) （30972269）
辽宁省创新团队项目(2007t017) （2007t017）
"百千万人才工程"资助项目(2008921059). （2008921059）
