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学科主题: Biotechnology & Applied Microbiology; Marine & Freshwater Biology
题名: Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp
作者: Jang, Songhun1, 2; Liu, Hang1, 3; Su, Jianguo1; Dong, Feng1, 3; Xiong, Feng1, 3; Liao, Lanjie1; Wang, Yaping1; Zhu, Zuoyan1
通讯作者: Wang, YP, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China
关键词: Bacterial artificial chromosome ; Grass carp ; Gene-specific hybridization
刊名: MARINE BIOTECHNOLOGY
发表日期: 2010-06-01
DOI: 10.1007/s10126-010-9268-0
卷: 12, 期:3, 页:261-266
收录类别: SCI
文章类型: Article
部门归属: [Jang, Songhun; Liu, Hang; Su, Jianguo; Dong, Feng; Xiong, Feng; Liao, Lanjie; Wang, Yaping; Zhu, Zuoyan] Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China; [Jang, Songhun] Kim IL Sung Univ, Coll Life Sci, Pyongyang, North Korea; [Liu, Hang; Dong, Feng; Xiong, Feng] Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: Development Plan of the State Key Fundamental Research of China [2009CB118701]; National Natural Science Foundation of China [30540084]
类目[WOS]: Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
研究领域[WOS]: Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
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.
关键词[WOS]: CATFISH ICTALURUS-PUNCTATUS ; TOLL-LIKE RECEPTORS ; PHYSICAL MAP ; BAC LIBRARY ; PARALICHTHYS-OLIVACEUS ; JAPANESE FLOUNDER ; ATLANTIC SALMON ; RAINBOW-TROUT ; GENOME ; EVOLUTION
语种: 英语
WOS记录号: WOS:000278095400003
ISSN号: 1436-2228
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/13611
Appears in Collections:鱼类生物学及渔业生物技术研究中心_期刊论文

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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

Recommended Citation:
Jang, Songhun; Liu, Hang; Su, Jianguo; Dong, Feng; Xiong, Feng; Liao, Lanjie; Wang, Yaping; Zhu, Zuoyan.Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Grass Carp,MARINE BIOTECHNOLOGY,2010,12(3):261-266
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