位置:成果数据库 > 期刊 > 期刊详情页
家蚕微孢子虫(Nosema bombycis)染色体DNA的分离纯化
  • ISSN号:0257-4799
  • 期刊名称:蚕业科学
  • 时间:2012
  • 页码:523-528
  • 分类:S884.21[农业科学—特种经济动物饲养;农业科学—畜牧兽医]
  • 作者机构:[1]西南大学,家蚕基因组生物学国家重点实验室,重庆400716, [2]重庆师范大学动物生物学重点实验室,重庆400047
  • 相关基金:国家自然科学基金重点项目(No.30930067); 重庆市科技攻关重大项目(No.2006AA5019)
  • 相关项目:家蚕微孢子虫孢壁蛋白质组及其在侵染中的作用研究
中文摘要:

为了获取可供家蚕微孢子虫(Nosema bombycis)单染色体测序的基础材料,通过脉冲场凝胶电泳(PFGE)技术分离家蚕微孢子虫CQ1株系的染色体DNA,随后切取含有目的染色体DNA条带的琼脂糖胶块,分别利用电洗脱法、柱式胶回收试剂盒法及玻璃奶胶回收试剂盒法进行染色体DNA的分离纯化实验。结果显示3种方法均能回收到目的染色体DNA,但分离纯化的效率和样品质量存在差异:电洗脱法对胶块中的DNA损失较大,回收的染色体DNA含量低,而柱式胶回收试剂盒回收的染色体DNA断裂严重,因此这2种方法获得的样品质量均达不到染色体建库测序的要求;通过优化后的玻璃奶胶回收试剂盒法获得的染色体DNA相对完整且污染较少,质量检验分析133μL样品中的DNA质量浓度为8.495 ng/μL(DNA总量1.13μg),达到后续构建染色体测序文库的要求。玻璃奶胶回收试剂盒法不仅适用于家蚕微孢子虫染色体DNA的分离纯化,也可供基因组较小的物种制备单染色体建库测序材料参考。

英文摘要:

To obtain basic materials for Nosema bombycis single chromosome sequencing, in this study, pulsed field gel electrophoresis (PFGE) was used to isolate chromosomal DNA of N. bombycis CQ1 strain. Agarose gels containing the target chromosomal DNA band were excised and subjected to further isolation and purification by means of electroelution, column gel recovery kit and glassmilk gel recovery kit. The results showed that all three methods could recover the target DNA but differed in efficiency of isolation and purification and in quality of obtained sample. Electroelution method led to high loss of DNA in the gel and the content of chromosomal DNA was low, while large portion of chromosomal DNA recovered by column gel extraction kit was in small pieces. The quality of sample purified using these two methods could not satisfy the sequencing requirement for library construction of chromosome. The modified glassmilk gel recovery kit method could recover relatively complete and less polluted chromosomal DNA. Quality inspection indicated that the mass concentration of sample DNA was 8. 495 ng/μL (the total DNA quantity of 133μL sample was 1. 13μg), and the sample reached the requirement of subsequent chromosome sequencing and library construction. The glassmilk gel recovery kit method is not only applicable to isolate and purify chromosomal DNA of N. bombycis, it can also be referred in prepara-tion of sequencing material for library construction of single chromosome of species with small genome.

同期刊论文项目
同项目期刊论文
期刊信息
  • 《蚕业科学》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国蚕学会 中国农业科学院蚕业研究所
  • 主编:郭锡杰
  • 地址:江苏省镇江市四摆渡中国农业科学院蚕业研究所
  • 邮编:212018
  • 邮箱:CYKE@chinajournal.net.cn
  • 电话:0511-85616835
  • 国际标准刊号:ISSN:0257-4799
  • 国内统一刊号:ISSN:32-1115/S
  • 邮发代号:28-23
  • 获奖情况:
  • 中国科协优秀科技期刊,全国农业学会优秀期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:7742