摘要
利用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,以高含油量蓖麻品种油蓖5号开花后第15d胚和36d胚为实验材料构建36d成熟期胚消减cDNA文库。文库的插入片段平均长度约400bp,随机挑取700个克隆进行PCR筛选,获得596个阳性克隆,经过点杂交筛选后选择了521个阳性克隆进行测序。经过同源性比对归并后得到96个差异表达基因,其中包括31个未知基因。已知基因涉及油脂合成、糖的分解、蛋白质贮藏与降解等多个方面。本研究还通过RT-PCR检测了部分可能与蓖麻油脂合成相关基因的差异表达水平,对其可能的功能进行了简要的分析。
The 15 day embryos and the 36 day embryos of high oil content variety Youbi 5 were used to construct suppression subtractive cDNA library of 36 day embryo by suppression subtractive hybridization （SSH）. The inserted fragment sizes were 400 bp on average, and 700 clones were verified by PCR, among which 596 clones were used to be performed by dot blot hybridization. After dot blot hybridization, 521 clones were obtained and sequenced. 96 differentially expressed genes which including 31 unknown functional genes, were obtained by homologous BLAST, Other genes were known to be related to oil synthesis, sugar degradation, protein storing and degradation, etc. Parts of differentially expressed genes probably relating to oil synthesis were detected by RT - PCR, and their functions were also analysed and discussed.
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2009年第3期305-310,共6页
Chinese Journal of Oil Crop Sciences
基金
中国农业科学院油料作物研究所所长基金
