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

[目的]研究传染性皮下及造血组织坏死病毒（Infectious hypodermal andhematopoietic necrosis virus,IHH-NV）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...

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Bibliographic Details
Published in南方农业学报 Vol. 44; no. 11; pp. 1899 - 1903
Main Author 曾地刚 陈秀荔 谢达祥 赵永贞 黎铭 陈晓汉
Format Journal Article
LanguageChinese
Published 广西水产科学研究院,南宁,530021 2013
Subjects
IHHNV
凡纳滨对虾
差异表达基因
转录本数目（RPKM）
高通量测序技术
差异表达基因
高通量测序技术
RPKM
IHHNV
differentially expressed genes
Litopenaeus vannamei
凡纳滨对虾
转录本数目(RPKM)
high-throughput sequencing
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ISSN2095-1191
DOI10.3969/j:issn.2095-1191.2013.11.1899

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Summary:[目的]研究传染性皮下及造血组织坏死病毒（Infectious hypodermal andhematopoietic necrosis virus,IHH-NV）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tmigenes;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.
Bibliography:45-1381/S
Litopenaeus vannamei;IHHNV;differentially expressed genes;RPKM;high-throughput sequencing
ZENG Di-gang, CHEN Xiu-li, XIE Da-xiang, ZHAO Yong-zhen, LIMing, CHEN Xiao-han （Guangxi Institute of Fisheries, Nanning 530021, China）
[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 f
ISSN:2095-1191
DOI:10.3969/j:issn.2095-1191.2013.11.1899