Construction and analysis of a suppression subtractive hybridization (SSH) library of genic multiple-allele inherited male-sterility in Chinese cabbage (Brassica campestris L. ssp. pekinensis)
Construction and analysis of a suppression subtractive hybridization (SSH) library of genic multiple-allele inherited male-sterility in Chinese cabbage (Brassica campestris L. ssp. pekinensis)
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大白菜(Brassica Campestris L. ssp. pekinensis)基因多等位基因遗传雄性不育抑制消减杂交(SSH)文库的构建与分析
DOI：
10.5897/ajb10.2048
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发表时间：
2011-06
影响因子：
--
通讯作者：
Ji RQ, Xin XF, Li ZQ, Feng H
中科院分区：
文献类型：
--
作者：
Ji RQ, Xin XF, Li ZQ, Feng H
利用雄性不育性是生产杂交大白菜的关键方法。学报(皱眉头。)奥尔森。本研究采用抑制减法杂交（suppression subtractive hybridization， SSH）技术，构建了A/B系‘AB01’可育芽和不育芽间差异表达的cDNA文库。采用聚合酶链反应扩增（PCR）随机筛选阳性克隆，得到25个高质量序列（22个来自育性试验库，3个来自无菌试验库）。片段长度从77 ~ 469 bp不等。利用5个表达序列标签（est）对可育芽和不育芽的差异表达模式进行了筛选和验证。结果表明，3条est仅在可育芽中表达，2条est在不育芽中表达下调。经BLAST （Basic Local Alignment Search Tool）筛选和功能注释，25条ESTs序列与NCBI （National Center for Biotechnology Information）数据库中的已知序列同源。这些基因与已知的蛋白如花/芽发育蛋白、代谢相关蛋白、细胞结构蛋白、细胞生长/分裂蛋白和次级代谢相关蛋白具有同源性。结果表明，这些蛋白在白菜基因多等位基因遗传性雄性不育的核雄性不育进程中起着关键作用。
Utilization of male sterility is a key method for producing crossbred Chinese cabbage (Brassica rapa L. ssp. pekinensis (Lour.) Olsson. In this study, suppression subtractive hybridization (SSH) was used to construct sterility and fertility cDNA libraries, which included differentially, expressed clones between fertile and sterile buds of the A/B line ‘AB01’. The positive clones were randomly selected by polymerase chain reaction amplification (PCR) and 25 high quality sequences (22 from the fertile-tester library and three from the sterile-tester libraries) were generated. The fragment lengths varied from 77 to 469 bp. Differential expression patterns between fertile and sterile buds were selected and verified using five expressed sequence tags (ESTs). Results indicated that, three ESTs were expressed only in fertile buds and two ESTs were down-regulated in sterile buds. According to the Basic Local Alignment Search Tool (BLAST) screening and functional annotation, the 25 ESTs were homologous to known sequences deposited in National Center for Biotechnology Information (NCBI). These genes had homology to known proteins such as flowers/buds development proteins, metabolic-related proteins, cell structure proteins, cell growth/division proteins and secondary metabolic-related proteins. The results suggested that, these proteins played a critical role in nuclear male sterility progression of genic multiple-allele inherited male-sterility in Chinese cabbage.
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DOI：
10.1159/000099415
发表时间：
2007-02
期刊：
--
影响因子：
--
作者：
J. Hulshof
通讯作者：
J. Hulshof
DOI：
--
发表时间：
2008
期刊：
--
影响因子：
--
作者：
Tian Zi-hua
通讯作者：
Tian Zi-hua
影响因子：
--
作者：
Le-cheng Liu;Jiashu Cao;Xiaolin Yu;X. Xiang;Y. Fei
通讯作者：
Le-cheng Liu;Jiashu Cao;Xiaolin Yu;X. Xiang;Y. Fei
影响因子：
2.9
作者：
S.-H. Chen;Nien-June Chung;Y.-N. Wang;Clifford Y. L. Lee;Y.-L. Lee;Po-Fang Tsai
通讯作者：
S.-H. Chen;Nien-June Chung;Y.-N. Wang;Clifford Y. L. Lee;Y.-L. Lee;Po-Fang Tsai
DOI：
--
发表时间：
2007
期刊：
--
影响因子：
--
作者：
Xu Wei
通讯作者：
Xu Wei