甘蓝基因组细菌人工染色体文库的构建

doi:10.7668/hbnxb.2014.05.005

华北农学报 ›› 2014, Vol. 29 ›› Issue (5): 23-28. doi: 10.7668/hbnxb.2014.05.005

所属专题： 生物技术；蔬菜专题；

甘蓝基因组细菌人工染色体文库的构建

邢苗苗^1,2, 田多成², 李海龙², 姚丽君³, 冀瑞琴¹, 康俊根²

1. 沈阳农业大学园艺学院, 辽宁沈阳 110866;
2. 北京市农林科学院蔬菜研究中心, 农业部华北地区园艺作物生物学与种质创制重点实验室, 北京 100097;
3. 河北农业大学园艺学院, 河北保定 071001

收稿日期:2014-03-10 出版日期:2014-10-28
作者简介:邢苗苗(1989-), 女, 山西运城人, 在读硕士, 主要从事蔬菜遗传育种与分子生物学研究。
基金资助:
国家自然科学基金项目(31171958);“十二五”国家科技支撑计划项目(2012BAD02B01;2012BAD50G01);国家高技术研究发展计划项目“863”项目(2012AA100103;2012AA100105;2012AA020103)

Construction of a Bacterial Artificial Chromosome Library from Brassica oleracea

XING Miao-miao^1,2, TIAN Duo-cheng², LI Hai-long², YAO Li-jun³, JI Rui-qin¹, KANG Jun-gen²

1. College of Horticulture, Shenyang Agricultural University, Shenyang 110866, China;
2. Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(North China), Ministry of Agriculture, Beijing 100097, China;
3. College of Horticulture, Agricultural University of Hebei, Baoding 071001, China

Received:2014-03-10 Published:2014-10-28

摘要/Abstract

摘要： 细菌人工染色体(BAC)文库是克隆甘蓝抗病优质重要基因的基础,以抗枯萎病、富含硫代葡萄糖苷的结球甘蓝自交系R4P1为材料,美国Epicentre公司的CopyControl^TM pCC1BAC^TM为载体,构建了甘蓝细菌人工染色体文库。该文库有73 344个克隆,插入片段平均大小约97 kb,空载率<3%,覆盖甘蓝基因组约10.9倍,筛选到甘蓝任一基因的概率为99.99%,这些结果表明,构建的文库质量较好,可用于后续分析。构建的甘蓝BAC文库不仅可用于枯萎病基因的克隆,而且为克隆其他重要的功能基因及基因组学等的研究奠定了基础。

关键词: 甘蓝, 细菌人工染色体文库, 枯萎病基因, 克隆

Abstract: Bacterial artificial chromosome library is the foundation of important and high quality disease-resistant gene cloning from Brassica oleracea.A bacterial artificial chromosome library was constructed from R4P1,a Brassica oleracea varieties highly resisting to fusarium wilt and being in rich of glucosinolates,with the vector CopyControl^TM pCC1BAC^TM.The library consists of 73 344 clones with an average insert size of about 97 kb,and in which the empty clones are under 3%.Thus,the library provides 10.9 haploid genome equivalents,the probability of screening to any gene of B.oleracea is 99.99%,these results indicate that this is a high quality BAC library suitable for the following analysis.The BAC library of B.oleracea could not only be used for the clone of resistant genes to fusarium wilt,but also lay foundations for isolating other interesting genes and research the genomics of B.oleracea,et al.

Key words: Brassica oleracea, Bacterial artificial chromosome library, Fusarium wilt resistant gene, Clone

中图分类号:

邢苗苗, 田多成, 李海龙, 姚丽君, 冀瑞琴, 康俊根. 甘蓝基因组细菌人工染色体文库的构建[J]. 华北农学报, 2014, 29(5): 23-28. doi: 10.7668/hbnxb.2014.05.005.

XING Miao-miao, TIAN Duo-cheng, LI Hai-long, YAO Li-jun, JI Rui-qin, KANG Jun-gen. Construction of a Bacterial Artificial Chromosome Library from Brassica oleracea[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(5): 23-28. doi: 10.7668/hbnxb.2014.05.005.

http://www.hbnxb.net/CN/Y2014/V29/I5/23

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