Construction of cDNA library from intestine, mesentery and coelomocyte of Apostichopus japonicus Selenka infected with Vibrio sp and a preliminary analysis of immunity-related genes

doi:10.1007/s11802-012-1863-4

CORC > 自然资源部第一海洋研究所 > 自然资源部第一海洋研究所 > 业务部门 > 海洋生态研究中心

	Construction of cDNA library from intestine, mesentery and coelomocyte of Apostichopus japonicus Selenka infected with Vibrio sp and a preliminary analysis of immunity-related genes
	Liu Hongzhan 2; Zheng Fengrong 1; Sun Xiuqin 1; Cai Yimei 3
刊名	JOURNAL OF OCEAN UNIVERSITY OF CHINA
	2012-06
卷号	11 期号:2 页码:187-196
关键词	Apostichopus japonicus cDNA library expressed sequence tags immunity-related genes real-time PCR
ISSN号	1672-5182
DOI	10.1007/s11802-012-1863-4
英文摘要	The aquaculture of sea cucumber Apostichopus japonicus (Echinodermata, Holothuroidea) has grown rapidly during recent years and has become an important sector of the marine industry in Northern China. However, with the rapid growth of the industry and the use of non-standard culture techniques, epidemic diseases of A. japonicus now pose increasing problems to the industry. To screen the genes with stress response to bacterial infection in sea cucumber at a genome wide level, we constructed a cDNA library from A. japonicus Selenka (Aspidochirotida: Stichopodidae) after infecting them with Vibrio sp. for 48 h. Total RNA was extracted from the intestine, mesentery and coelomocyte of infected sea cucumber using Trizol and mRNA was isolated by Oligotex mRNA Kits. The ligated cDNAs were transformed into DH5 alpha, and a library of 3.24x10(5) clones (3.24x10(5) cfu mL(-1)) was obtained with the sizes of inserted fragments ranging from 0.8 to 2.5 kb. Sequencing the cDNA clones resulted in a total of 1106 ESTs that passed the quality control. BlastX and BlastN searches have identified 168 (31.5%) ESTs sharing significant homology with known sequences in NCBI protein or nucleotide databases. Among a panel of 25 putative immunity-related genes, serum lectin isoform, complement component 3, complement component 3-like genes were further studied by real-time PCR and they all increased more than 5 fold in response to Vibrio sp. challenge. Our library provides a valuable molecular tool for future study of invertebrate immunity against bacterial infection and our gene expression data indicates the importance of the immune system in the evolution and development of sea cucumber.
资助项目	Key Project of Youth Marine Science funded for Oceanography[2009119]
WOS研究方向	Oceanography
语种	英语
出版者	OCEAN UNIV CHINA
WOS记录号	WOS:000303526400011
内容类型	期刊论文
源URL	[http://ir.fio.com.cn/handle/2SI8HI0U/4105]
专题	业务部门_海洋生态研究中心
作者单位	1.SOA, Inst Oceanog 1, Qingdao 266061, Peoples R China; 2.Ocean Univ China, Coll Marine Life Sci, Qingdao 266003, Peoples R China; 3.Chinese Acad Sci, Beijing Inst Genom, Beijing 100029, Peoples R China
推荐引用方式 GB/T 7714	Liu Hongzhan,Zheng Fengrong,Sun Xiuqin,et al. Construction of cDNA library from intestine, mesentery and coelomocyte of Apostichopus japonicus Selenka infected with Vibrio sp and a preliminary analysis of immunity-related genes[J]. JOURNAL OF OCEAN UNIVERSITY OF CHINA,2012,11(2):187-196.
APA	Liu Hongzhan,Zheng Fengrong,Sun Xiuqin,&Cai Yimei.(2012).Construction of cDNA library from intestine, mesentery and coelomocyte of Apostichopus japonicus Selenka infected with Vibrio sp and a preliminary analysis of immunity-related genes.JOURNAL OF OCEAN UNIVERSITY OF CHINA,11(2),187-196.
MLA	Liu Hongzhan,et al."Construction of cDNA library from intestine, mesentery and coelomocyte of Apostichopus japonicus Selenka infected with Vibrio sp and a preliminary analysis of immunity-related genes".JOURNAL OF OCEAN UNIVERSITY OF CHINA 11.2(2012):187-196.