Molecular cloning and characterization of a novel pollen predominantly membrane protein gene BcMF12 from Brassica campestris ssp. chinensis
Molecular cloning and characterization of a novel pollen predominantly membrane protein gene BcMF12 from Brassica campestris ssp. chinensis
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DOI：
10.1007/s11033-009-9449-y
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发表时间：
2009-01
影响因子：
2.8
通讯作者：
Jianghua Song;Lixin Zhang;Jiashu Cao
中科院分区：
文献类型：
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作者：
Jianghua Song;Lixin Zhang;Jiashu Cao
从大白菜（Brassica campestrisL.）SSP. chinensisMakino）的花粉特异性cDNA片段（DN 237936）。该cDNA长度为1,155 bp，开放阅读框为894 bp，能够编码297个氨基酸的推定多肽，估计分子量为34.6 kDa，预测等电点为9.6。比较和生物信息学分析表明，BcMF 12与公共数据库中已发表的一些膜蛋白序列高度相似，包含6个高度保守的跨膜结构域，对应6个高度疏水的区域。这表明BcMF 12可能是一种膜蛋白。RNA凝胶印迹分析表明，BcMF 12的转录本在花蕾、花和花药中含量丰富，而在根、茎、叶和雌蕊中未检测到。此外，BcMF 12转录本在花粉发育的后期也可检测到。对转BcMF 12反义基因植株的花粉形态观察表明，转基因植株的花粉粒大多数是异常的。这些结果表明BcMF 12是一种新的花粉优先膜蛋白，在大白菜花粉发育过程中起重要作用。
A novel membrane protein gene,BcMF12, was isolated from Chinese cabbage (Brassica campestrisL. ssp. chinensisMakino) using rapid amplification of the cDNA ends based on a pollen-specific cDNA fragment (DN237936). The cDNA was 1,155 bp in length with an open reading frame of 894 bp capable of encoding a putative polypeptide of 297 amino acids with an estimated molecular mass of 34.6 kDa and a predicted isoelectric point of 9.6. Comparative and bioinformatics analyses revealed thatBcMF12showed high similarities with some membrane protein sequences previously published in the public database and contained six highly conserved transmembrane domains corresponding to six highly hydrophobic regions. This indicates thatBcMF12may be a putative membrane protein. RNA gel blot analysis indicated that the transcripts ofBcMF12were abundant in the flower bud, flower and anther, but not detected in the root, stem, leaf and pistil. Moreover, theBcMF12transcripts were detectable at the late stages of pollen development. Morphological investigations of pollen from theBcMF12antisense transgenic plants showed that most of pollen grains of transgenic plants were abnormal. These results strongly suggest that BcMF12 is a novel pollen-preferentially membrane protein which play an important role during the pollen development in Chinese cabbage.