期刊论文

中文

中国农业科学

0578-1752

2013

46

1

9-17

10.3864/j.issn.0578-1752.2013.01.002

国家“863”计划项目（2012AA101105-4）； 国家“十二五”科技支撑计划（2012BAD03B01-2）； 国家现代农业产业技术体系建设专项资金（CARS-05）；

本校为第一机构

【目的】从大麦BAC文库中快速筛选包含目的基因HvGW2的阳性BAC克隆。【方法】构建BAC文库三维混合池,设计2对HvGW2特异性引物,进行BAC文库筛选,对含有目的基因HvGW2的阳性BAC克隆进行测序,获得编码大麦GW2蛋白的基因组DNA序列,并与禾本科中的GW2进行同源进化分析。【结果】构建了大麦BAC文库的三维混合池160个,文库空载率为0.18%;筛选到含HvGW2的阳性BAC克隆3个,其中,BAC克隆196M02包含完整编码该基因的序列;HvGW2包含8个外显子和7个内含子,具有与其它植物GW2相似的结构特征和保守的功能结构域;同源进化分析结果显示,大麦与小麦的GW2蛋白同源性最高;序列分析发现,HvGW2第6内含子上包含2个不同类...

[Objective] The objective of this study was to isolate HvGW2 gene from barley BAC library rapidly. [Method] Three-dimensional （3D） BAC pools were constructed and screened by using barley GW2 gene-specific PCR primers. Positive BAC clones were screened out and further sequenced, the obtained HvGW2 gene was used for homologous evolutionary analysis with those of other grasses. [Result] One hundred and sixty BAC DNA pools were prepared by 3D pooling strategy from barley BAC library with only 0.18% of empty clones. PCR-based screening demonstrate...