Li Yanyan, Cao Jiashu, Huang Li, Yu Xiaolin, Xiang Xun
Lab of Cell & Molecular Biology, Institute of Vegetable Science, Zhejiang University, 282 Kaixuan Road, Hangzhou, 310029, China.
Mol Biol Rep. 2008 Jun;35(2):207-14. doi: 10.1007/s11033-007-9072-8. Epub 2007 Mar 27.
A transcript-derived fragment (GenBank accession number DN237920.1) accumulated in the wild-type flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) was isolated and further investigated. The full length DNA and cDNA of the fragment were cloned by rapid amplification of cDNA ends. The gene, BcMF13, encodes a protein of 73 amino acids and is interrupted by an intron of 106 bp (GenBank accession number EF158459). Southern blot analysis revealed that BcMF13 could be a single-copy gene in the Chinese cabbage genome. Sequence blast analysis showed that BcMF13 was a new gene. In EST database, those sequences share 96-98% identity with BcMF13 cDNA all came from flower buds, microspores, anthers of Brassica, which proved BcMF13 homologs closely related to the development of male gametogenesis in Brassica. RT-PCR discovered that it is exclusively expressed in stage four and five flower buds of fertile line, strongly expressed in stamens. Successful suppression of BcMF13 gene expression by RNA antisense strategy greatly reduced the normal pollen grains, suggesting that BcMF13 was essential in pollen development in Brassica.
从大白菜(Brassica campestris L. ssp. chinensis Makino,同义名B. rapa ssp. chinensis)的野生型花蕾中分离出一个转录本衍生片段(GenBank登录号DN237920.1)并进行了进一步研究。通过cDNA末端快速扩增克隆了该片段的全长DNA和cDNA。BcMF13基因编码一个73个氨基酸的蛋白质,被一个106 bp的内含子打断(GenBank登录号EF158459)。Southern杂交分析表明BcMF13可能是大白菜基因组中的单拷贝基因。序列比对分析表明BcMF13是一个新基因。在EST数据库中,那些与BcMF13 cDNA具有96 - 98%同一性的序列均来自芸苔属植物的花蕾、小孢子、花药,这证明BcMF13同源物与芸苔属植物雄配子体发育密切相关。RT-PCR发现它仅在可育系的四期和五期花蕾中表达,在雄蕊中强烈表达。通过RNA反义策略成功抑制BcMF13基因表达极大地减少了正常花粉粒,表明BcMF13在芸苔属植物花粉发育中至关重要。