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甜瓜(Cucumis melo L.)果实成熟过程中表达的两种mRNA的cDNA克隆的鉴定

Characterization of two cDNA clones for mRNAs expressed during ripening of melon (Cucumis melo L.) fruits.

作者信息

Aggelis A, John I, Karvouni Z, Grierson D

机构信息

Department of Physiology and Environmental Science, The University of Nottingham, Loughborough, UK.

出版信息

Plant Mol Biol. 1997 Jan;33(2):313-22. doi: 10.1023/a:1005701730598.

Abstract

In vitro translation of mRNAs and polyacrylamide gel electrophoresis of proteins from melons revealed that several mRNAs increased in amount during ripening, indicating the existence of other ripening genes in addition to those cloned previously. To identify ripening-related genes we have screened a ripe melon cDNA library and isolated two novel cDNA clones (MEL2 and MEL7) encoding unidentified proteins. Southern analysis revealed that MEL2 and MEL7 are encoded by low-copy-number genes. The MEL2 cDNA clone is near full-length, corresponds to a 1600 nucleotide mRNA that accumulates during ripening and encodes a predicted protein rich in hydrophobic amino acids. The MEL7 cDNA clone is full-length, corresponds to a mRNA of 0.7 kb which accumulates during early ripening stages and is also present at low levels in other organs of the melon plant. The MEL7 predicted polypeptide is 17 kDa and shows significant homology with the major latex protein from opium-poppy. Wounding and ethylene treatment of unripe melon fruits 20 days after anthesis showed that MEL2 and MEL7 mRNAs are only induced by ethylene.

摘要

对甜瓜的mRNA进行体外翻译以及对蛋白质进行聚丙烯酰胺凝胶电泳分析发现,在成熟过程中,有几种mRNA的量增加了,这表明除了先前克隆的那些基因外,还存在其他成熟基因。为了鉴定与成熟相关的基因,我们筛选了一个成熟甜瓜的cDNA文库,并分离出两个编码未知蛋白质的新cDNA克隆(MEL2和MEL7)。Southern分析表明,MEL2和MEL7由低拷贝数基因编码。MEL2 cDNA克隆接近全长,对应于一个1600个核苷酸的mRNA,该mRNA在成熟过程中积累,并编码一种预测的富含疏水氨基酸的蛋白质。MEL7 cDNA克隆是全长的,对应于一个0.7 kb的mRNA,该mRNA在成熟早期阶段积累,并且在甜瓜植株的其他器官中也以低水平存在。MEL7预测的多肽为17 kDa,与罂粟中的主要乳胶蛋白具有显著的同源性。对开花后20天的未成熟甜瓜果实进行创伤和乙烯处理表明,MEL2和MEL7 mRNA仅由乙烯诱导。

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