利用高通量测序技术分析IHHNV感染凡纳滨对虾的基因差异表达  

Analysis on differential gene expression in shrimp Litopenaeus vannamei induced by IHHNV infection using high-throughput sequencing

作　　者：曾地刚 陈秀荔 谢达祥 赵永贞 黎铭 陈晓汉 

机构地区：[1]广西水产科学研究院,南宁530021

出　　处：《南方农业学报》2013年第11期1899-1903,共5页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31160531)

摘　　要：【目的】研究传染性皮下及造血组织坏死病毒(Infectious hypodermal andhematopoietic necrosis virus,IHHNV)感染凡纳滨对虾的基因差异表达,为进一步了解IHHNV与凡纳滨对虾的相互作用分子机制提供参考。【方法】采用454高通量测序技术对IHHNV感染和对照凡纳滨对虾进行转录组测序,测序获得的序列除去接头后用iAssembler软件进行拼接得到unigene(单一基因序列),通过计算各unigene的转录本数目(RPKM)确定差异表达基因,并以实时荧光定量PCR验证基因的差异表达。【结果】采用454高通量测序技术对IHHNV感染和对照凡纳滨对虾进行转录组测序,共获得208690条短序列,序列拼接共得到21912条unigenes;通过对IHHNV感染和对照凡纳滨对虾的测序数据比较分析,结果发现221个差异表达基因,包括81个上调表达基因和140个下调表达基因。经实时荧光定量PCR验证,454高通量测序数据统计的基因表达差异结果可靠。【结论】IHHNV感染显著影响凡纳滨对虾免疫相关基因的表达,而利用高通量测序技术可有效检测出这些差异表达基因。[Objective]The differential gene expression in shrimp Litopenaeus vannamei induced by IHHNV infection was studied in order to provide references for further understanding the molecular interaction mechanisms of IHHNVshrimp interaction.[Method]The transcriptomes of the IHHNV-infected and control Litopenaeus vannamei were sequenced using the 454 high-throughput sequencing technology.After the removal of adaptors and sequence assembly by using the iAssembler software,unigenes （unique sequences） were obtained.Differentially expressed genes were determined by calculating the transcripts of each unigene （RPKM）,and were validated by qRT-PCR.[Result]The sequencing produced a total of 208,690 short sequences,and the assembly generated 21,912 unigenes.Comparison between the sequencing data from IHHNV-infected and control Litopenaeus vannamei revealed 221 differentially expressed genes,including 81 upregulated genes and 140 down-regulated genes.qRT-PCR validation verified that the statistical gene expression results from 454 high-throughput sequencing data were realiable.[Conclusion]IHHNV infection had significant impact on immunerelated gene expression in Litopenaeus vannamei,while these differentially expressed genes could he detected using highthroughput sequencing.

关 键 词：凡纳滨对虾 IHHNV 差异表达基因 转录本数目(RPKM) 高通量测序技术 

分 类 号：S945.46[农业科学—水产养殖]