巴西橡胶树环锌指蛋白基因克隆及其分子特性(英文)

Cloning and Molecular Characterization of a RING Zinc-Finger Gene of Hevea brasiliensis

在先前的研究中通过抑制缩减杂交获得了一个在巴西橡胶树胶乳中特异表达的片段(HbSSH10),该片段含有“RINGfinger”或“C3HC4”保守序列。根据HbSSH10的序列信息设计引物并通过3'-RACE和5'-RACE的方法,获得了一个全长的cDNA(HbRZF)。该cDNA含有589个核苷酸,含有完整的阅读框架,编码156个氨基酸。从它推导出的氨基酸序列中含有“RINGfinger”或“C3HC4”保守区(氨基酸100￣144)。该氨基酸序列与Poncirustrifoliata、Arabidopsisthaliana和Thellungiellahalophila的环锌指蛋白的同源性分别为48%、52%和50%。Northern杂交分析表明HbRZF在胶乳中大量表达,在叶片中微量表达,而在根和花中几乎没有表达。茉莉酸处理可以诱导胶乳中HbRZF的表达,而乙烯对胶乳中HbRZF的表达基本上没有影响。

Using the information about the sequence from a differentially expressed clone （designated as HbSSH10） encodes a protein specifying a cysteine-rich sequence containing a putative ＂RING finger＂ or ＂C3HC4＂ consensus motif that was cloned recently by the subtractive hybridization between latex and leaves from rubber tree （Hevea brasiliensis）. A full-length cDNA encoding C3HC4 type zinc-finger protein was isolated and characterized from rubber tree. Sequence analysis revealed that the ORFs of HbRZF encode 156 amino acid residues with a total predicted molecular mass of 17.2 kD, HbRZF protein having a putative ＂RING finger＂ segment （amino acid residues 100-144）. The deduced amino acid sequences of HbRZF showed high identities of 48%, 52% and 50% to those of the ring zinc protein from Poncirus trifoliata, Arabidopsis thaliana, Thellungiella halophila. The result of Northern blot analysis indicated that the transcripts of the HbRZF were expressed more in the latex than in the leaves, whereas little expression was detected in roots and flowers. The transcription of HbRZF was induced by jasmonic acid, whereas ethylene had little effect.