脂多糖刺激的青石斑鱼脾脏中差异表达基因的鉴定。
Identification of differentially expressed genes in lipopolysaccharide-stimulated yellow grouper Epinephelus awoara spleen.
作者信息
Wang Li, Wu Xinzhong
机构信息
Laboratory of Marine Life Science and Technology, College of Animal Sciences, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang 310029, China.
出版信息
Fish Shellfish Immunol. 2007 Aug;23(2):354-63. doi: 10.1016/j.fsi.2006.11.009. Epub 2006 Nov 30.
A subtracted cDNA library from lipopolysaccharide (LPS)-stimulated Epinephelus awoara spleen was constructed using the suppression subtractive hybridization (SSH). Random clones (209) were selected and sequenced. After assembling, 36 contigs and 56 singlets (accession numbers: EB410743-EB410834) were finally obtained, some of which were immune-related genes. A reverse transcription-polymerase chain reaction (RT-PCR) analysis of the expression patterns of eight transcripts showed that seven of them were up-regulated after 24 h of LPS stimulation. Furthermore, full-length cDNAs of homologues of defender against cell death 1 (DAD1) and allograft inflammatory factor-1 (AIF-1) were obtained by RACE-PCR.
利用抑制性消减杂交（SSH）技术构建了脂多糖（LPS）刺激的青石斑鱼脾脏消减cDNA文库。随机挑选209个克隆进行测序。组装后，最终获得36个重叠群和56个单拷贝序列（登录号：EB410743 - EB410834），其中一些是免疫相关基因。对8个转录本的表达模式进行逆转录聚合酶链反应（RT-PCR）分析表明，其中7个在LPS刺激24小时后上调。此外，通过RACE-PCR获得了细胞死亡防御者1（DAD1）和同种异体移植炎症因子-1（AIF-1）同源物的全长cDNA。
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