副溶血弧菌刺激下拟穴青蟹转录组和表达谱分析及部分免疫基因的克隆-生物化学与分子生物学专业论文.docx

汕头大学 汕头大学 2013 硕士学位论文 PAGE PAGE VI Abstract The mud crab Scylla paramamosain is one of the most economically important mariculture in south China. In recent years, infectious diseases have prevented crab aquaculture from sustainable development with the expanding breeding scale and high density cultication. Because of the limited knownledge of S.paramamosain，s immune related genes and immune system, it is still at an early stage for disease prevention and control in mud crab aquaculture. In order to fully understand S.paramamosain，s immune system and be good for healthy crab breeding, this paper intended use high-throughput deep sequencing technology to investigate the transcriptome and the expression profiles of the mud crab S.paramamosain infected with Vibrio parahaemolyticus. The results would lay the basis for further research on the important immune related genes in S.paramamosain. The results were shown as follows: Transcriptome analysis in S.paramamosain during V.parahaemolyticus infection Approximately 52,934,042 clean reads were generated by using Solexa Hiseq2000 high-throughput deep sequencing for the transcriptome of S.paramamosain during V.parahaemolyticus infection. The total number of nucleotides was 4,764,063,780 bp. Clean reads were obtained and assembled into 186,193 contigs. Eventually we acquired 81,709 unigenes with a mean length of 656 bp from the transcriptome libraries. Blasted with the Nr, Swissprot, GO and COG databases, 23,978, 20,147, 10,566 and 9,135 transcripts were annotated, respectively. Approximately 25,349 transcripts might be involved in 258 metabolic or signalling pathways. Expression profiling analysis in S.paramamosain during V.parahaemolyticus infection Using gene expression profiling, we analyzed the expression profiles between V.parahaemolyticus-infected crabs and the control crabs, including 538 significantly up regulated genes and 675 significantly down regulated genes. 96 genes related to immune and signal t