2009•Zhongguo yufang shouyi xuebaoRequires access
Construction of cDNA SSH libraries for grunniens peripheral blood mononuclear cell.
He YanHua, Zhizhong Jing, Guohua Chen, Fang YongXiang, Meng Xue-lian, LI Xiao-qing, Duan Feng-yun, Shengfu Wang
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Abstract
SA subtractive cDNA library was constructed by suppression subtractive hybridization(SSH) using cDNA from Yak peripheral blood mononuclear cells(PBMC) treated with lipopolysaccharide(LPS) and concanavalinA(ConA).The library contained 93 % recombinants with insert size ranged from 200 bp to 1 000 bp.One hundred positive clones were randomly sequenced and analyzed,and 24 differentially expressed genes and 3 novel ESTs were identified.The subtraction efficiency of the SSH cDNA library was confirmed by randomly testing 6 non-repeat sequences by semi-quantitative RT-PCR,of which 5 differential expression molecules were detected.These results suggest that SSH is an effective technique to study gene differential expression gene.