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通过差异显示法鉴定棉花纤维特异性酰基载体蛋白cDNA

Identification of a cotton fiber-specific acyl carrier protein cDNA by differential display.

作者信息

Song P, Allen R D

机构信息

Department of Biological Sciences, Texas Tech University, Lubbock 79409, USA.

出版信息

Biochim Biophys Acta. 1997 Apr 10;1351(3):305-12. doi: 10.1016/s0167-4781(96)00218-7.

DOI:10.1016/s0167-4781(96)00218-7
PMID:9130594
Abstract

Transcripts from immature fibers and stripped ovules (fibers removed) of cotton (Gossypium hirsutum L.) were compared by differential display to identify cDNA fragments that represent mRNAs that are expressed primarily in cotton fibers. Eight independent fiber-specific cDNA fragments were isolated. One of these cDNAs had strong sequence similarity with acyl carrier protein (ACP). A full-length cDNA for the cotton fiber-specific ACP was isolated using a PCR cDNA library screening technique. This 713 bp cDNA has an open reading frame that encodes a 136 amino acid polypeptide. Overall nucleotide and amino acid sequence identities with other plant ACP gene sequences averaged 66% and 60% respectively. A 19 amino acid sequence surrounding the prosthetic group attachment site is nearly identical to other plant ACP genes. Northern blot analyses showed that transcripts homologous to this fiber-specific ACP cDNA were predominantly expressed during the elongation stage of fiber development. Initial genomic Southern blot analysis indicated that a single copy of the fiber-specific ACP gene may be present in both the cotton A and D genomes, since diploid Gossypium species with A or D genomes gave identical bands. We speculate that this putative fiber-specific ACP may play an important role in rapidly elongating cotton fibers by contributing to the synthesis of membrane lipids. It is also apparent that during the evolution of cotton a member of the ACP gene family has been recruited for specific expression in cotton fibers.

摘要

通过差异显示比较了棉花(陆地棉)未成熟纤维和去除胚珠(去除纤维)的转录本,以鉴定代表主要在棉花纤维中表达的mRNA的cDNA片段。分离出了八个独立的纤维特异性cDNA片段。其中一个cDNA与酰基载体蛋白(ACP)具有很强的序列相似性。使用PCR cDNA文库筛选技术分离出了棉花纤维特异性ACP的全长cDNA。这个713 bp的cDNA有一个开放阅读框,编码一个136个氨基酸的多肽。与其他植物ACP基因序列的总体核苷酸和氨基酸序列同一性分别平均为66%和60%。辅基附着位点周围的一个19个氨基酸的序列与其他植物ACP基因几乎相同。Northern印迹分析表明,与这个纤维特异性ACP cDNA同源的转录本在纤维发育的伸长阶段主要表达。最初的基因组Southern印迹分析表明,纤维特异性ACP基因的单拷贝可能存在于棉花A和D基因组中,因为具有A或D基因组的二倍体棉属物种给出了相同的条带。我们推测,这个假定的纤维特异性ACP可能通过促进膜脂的合成在快速伸长的棉花纤维中发挥重要作用。同样明显的是,在棉花进化过程中,ACP基因家族的一个成员已被招募用于在棉花纤维中特异性表达。

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