副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建

CORC > 集美大学

	副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建
	王淑红 ; 邹志华 ; 张子平 ; 施永春 ; 王艺磊
刊名	http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=twhx200803003&dbcode=CJFQ&dbname=CJFQ2008
	2012-06-05 ; 2012-06-05
关键词	cDNA文库均一化九孔鲍副溶血弧菌血细胞 cDNA library normalized Haliotis diversicolor supertexta Vibrio parahaemolyticus haemocytes S917.4
其他题名	Construction a normalized full-length cDNA library of haemocytes from Haliotis diversicolor supertexta infected with Vibrio parahaemolyticus
中文摘要	采用Trizol试剂提取副溶血弧菌感染的九孔鲍血细胞的总RNA,经Oligotex纯化得到mRNA.根据SMART技术原理合成双链cDNA.双链cDNA采用双链特异核酸酶进行cDNA的均一化,构建成九孔鲍血细胞的均一化全长cDNA文库.原始文库的库容为3.4×106cfu/cm3,重组率为92.3%,扩增后文库的滴度为2.6×1011cfu/cm3以上.从文库中随机挑取897个克隆,测序获得高质量ES-Ts814条,其中有23条Contigs;762条Singlets,Unigenes共785条,冗余率只有3.56%.以上结果说明所构建的文库质量好,完全可以满足后续基因克隆和表达序列标签测序工作的需要.; A normalized full-length haemocytes cDNA library was constructed in order to clone genes involving in Vibrio parahaemolyticus defenses from Haliotis diversicolor supertexta.Total RNAs were prepared from haemocytes using TRZOL reagent.Oligotex(QIAGEN) was used to separate the mRNA from total RNA.The "anchor first-strand cDNA" containing a symmetrical sfiI restriction enzyme sites(A&B) was synthesized by transcription of mRNA with the SMART technique.Double strand cDNA was normalized using Duplex-Specific Nuclease(DSN).The containing capacities of the unamplified cDNA library was 3.4×106 cfu/cm3 with a recombinant rate of 92.3% and that of amplified one was more than 1011 cfu/cm3.A total of 897 clones was random selected to be sequenced and 814 high quality expressed sequence tags were obtained.Among them,there were 23 Contigs and 762 Singlets,the redundancy was only 3.56%.The results indicated that a high quality library was meet the needs for the consequence gene clone and expressed sequence tag analysis.; 【作者单位】集美大学水产学院水产生物技术研究所福建省高校水产科学技术与食品安全重点实验室；德克萨斯州立大学化学和生化系；【作者英文名】WANG Shu-hong1,ZOU Zhi-hua1,ZHANG Zi-ping2,SHI Yong-chun1,WANG Yi-lei1(1.Fisheries College,Institute of Aquaculture Biotechnology,Key Laboratory of Scienceand Technology for Aquaculture and Food Safety,Fujian Province University,Jimei University,Xiam
语种	中文
内容类型	期刊论文
源URL	[http://ir.calis.edu.cn/hdl/235041/12221]
专题	集美大学
推荐引用方式 GB/T 7714	王淑红,邹志华,张子平,等. 副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建[J]. http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=twhx200803003&dbcode=CJFQ&dbname=CJFQ2008,2012, 2012.
APA	王淑红,邹志华,张子平,施永春,&王艺磊.(2012).副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建.http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=twhx200803003&dbcode=CJFQ&dbname=CJFQ2008.
MLA	王淑红,et al."副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建".http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=twhx200803003&dbcode=CJFQ&dbname=CJFQ2008 (2012).