Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp

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	Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp
	Jang, Songhun 1,2; Liu, Hang 1,3; Su, Jianguo 1; Dong, Feng 1,3; Xiong, Feng 1,3; Liao, Lanjie 1; Wang, Yaping 1; Zhu, Zuoyan 1
刊名	MARINE BIOTECHNOLOGY
	2010-06-01
卷号	12 期号:3 页码:261-266
关键词	Bacterial artificial chromosome Grass carp Gene-specific hybridization
ISSN号	1436-2228
通讯作者	Wang, YP, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China
中文摘要	Bacterial artificial chromosome (BAC) library is an important tool in genomic research. We constructed two libraries from the genomic DNA of grass carp (Ctenopharyngodon idellus) as a crucial part of the grass carp genome project. The libraries were constructed in the EcoRI and HindIII sites of the vector CopyControl pCC1BAC. The EcoRI library comprised 53,000 positive clones, and approximately 99.94% of the clones contained grass carp nuclear DNA inserts (average size, 139.7 kb) covering 7.4x haploid genome equivalents and 2% empty clones. Similarly, the HindIII library comprised 52,216 clones with approximately 99.82% probability of finding any genomic fragments containing single-copy genes; the average insert size was 121.5 kb with 2.8% insert-empty clones, thus providing genome coverage of 6.3x haploid genome equivalents of grass carp. We selected gene-specific probes for screening the target gene clones in the HindIII library. In all, we obtained 31 positive clones, which were identified for every gene, with an average of 6.2 BAC clones per gene probe. Thus, we succeeded in constructing the desired BAC libraries, which should provide an important foundation for future physical mapping and whole-genome sequencing in grass carp.
英文摘要	Bacterial artificial chromosome (BAC) library is an important tool in genomic research. We constructed two libraries from the genomic DNA of grass carp (Ctenopharyngodon idellus) as a crucial part of the grass carp genome project. The libraries were constructed in the EcoRI and HindIII sites of the vector CopyControl pCC1BAC. The EcoRI library comprised 53,000 positive clones, and approximately 99.94% of the clones contained grass carp nuclear DNA inserts (average size, 139.7 kb) covering 7.4x haploid genome equivalents and 2% empty clones. Similarly, the HindIII library comprised 52,216 clones with approximately 99.82% probability of finding any genomic fragments containing single-copy genes; the average insert size was 121.5 kb with 2.8% insert-empty clones, thus providing genome coverage of 6.3x haploid genome equivalents of grass carp. We selected gene-specific probes for screening the target gene clones in the HindIII library. In all, we obtained 31 positive clones, which were identified for every gene, with an average of 6.2 BAC clones per gene probe. Thus, we succeeded in constructing the desired BAC libraries, which should provide an important foundation for future physical mapping and whole-genome sequencing in grass carp.
学科主题	Biotechnology & Applied Microbiology; Marine & Freshwater Biology
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
研究领域[WOS]	Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
关键词[WOS]	CATFISH ICTALURUS-PUNCTATUS ; TOLL-LIKE RECEPTORS ; PHYSICAL MAP ; BAC LIBRARY ; PARALICHTHYS-OLIVACEUS ; JAPANESE FLOUNDER ; ATLANTIC SALMON ; RAINBOW-TROUT ; GENOME ; EVOLUTION
收录类别	SCI
资助信息	Development Plan of the State Key Fundamental Research of China [2009CB118701]; National Natural Science Foundation of China [30540084]
语种	英语
WOS记录号	WOS:000278095400003
公开日期	2010-12-23
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/342005/13611]
专题	水生生物研究所_鱼类生物学及渔业生物技术研究中心_期刊论文
作者单位	1.Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China 2.Kim IL Sung Univ, Coll Life Sci, Pyongyang, North Korea 3.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Jang, Songhun,Liu, Hang,Su, Jianguo,et al. Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp[J]. MARINE BIOTECHNOLOGY,2010,12(3):261-266.
APA	Jang, Songhun.,Liu, Hang.,Su, Jianguo.,Dong, Feng.,Xiong, Feng.,...&Zhu, Zuoyan.(2010).Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp.MARINE BIOTECHNOLOGY,12(3),261-266.
MLA	Jang, Songhun,et al."Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp".MARINE BIOTECHNOLOGY 12.3(2010):261-266.