Isolation and Characterization of the Third Gene Encoding a 16.9kDa Class I Low-molecular-mass Heat Shock Protein, Oshsp 16.9C, in Rice

水稻中第三個16.9kDa第一族低分子量熱休克蛋白質基因，Oshsp 16.9C，之分離與鑑定

張碧芳(Pi-Fang Linda Chang) ； 黃仲義(Chung-Yi Huang) ； 張法正(Fa-Cheng Chang) ； 曾冬筍(Tong-Seung Tseng) ； 林婉琦(Wan-Chi Lin) ； 林秋榮(Chu-Yung Lin)

《Botanical Bulletin of Academia Sinica》 42卷2期 (2001/04) Pp. 85-92

https://doi.org/10.7016/BBAS.200104.0085

摘要

利用水稻16.9kDa熱休克蛋白質(HSP)基因之cDNA，pTS1，作為南方墨點分析法之探針，可在以Eco RI限制切割之水稻基因組DNA樣品中，偵測到五個清晰的條帶，其大小分別為9.5、5.9、3.4、2.5及1.7kb，經過本研究室之分析，其中包含六個不同的基因。將5.9kb之DNA片段中可能包含一低分子量熱休克蛋白質基因。經由定序分析此3.5kb片段之核酸序列，推測其可能之轉譯產物為包含149個氨基酸之16.9kDa熱休克蛋白質，其等電點(PI)值估計為6.42。此基因和本研究室已發表的二個水稻第一族低分子量熱休克蛋白質基因，Oshsp 16.9A及Oshsp 16.9B，在轉譯區內之核酸序列具有相當高的同質性，分別有93.1%及94.3%的相同度。此基因之氨基酸推演序列和Oshp 16.9A及Oshsp 16.9B基因幾乎完全相同，分別只有11個及10個氨基酸之不同，因此將此基因定為Oshsp 16.9C(accession number:U81385)。已知水稻第一族低分子量熱休克蛋白質基因之3＇非轉譯區(3＇UTR)的核酸序列相似性較低，因此我們將Oshsp 16.9C之3＇非轉譯區作為探針，以分析其基因之表現情形。

關鍵字

熱休克蛋白質 ； 熱休克基因 ； 低分子量熱休克蛋白質 ； 水稻

並列摘要

Using rice cDNA pTS1 encoding a 16.9kDa heat shock protein (HSP) as a probe for Southern hybridization analysis, we observed five prominent bands of 9.5, 5.9, 3.4, 2.5, and 1.7kb in the Eco RI digests of rice genomic DNA and found them to contain six individual genes. The 5.9kb DNA fragment was further digested with Hind III to generate three fragments of 3.5, 1.7 and 0.7kb, and we found, using pTS1 cDNA as a probe, the 3.5kb fragment contained a putative low-molecular-mass (LMM) HSP gene. The DNA sequencing of 3.5kb fragment revealed encoding of a presumptive 16.9kDa HSP (149 amino acid residues) with predicted pI value of 6.42. The nucleotide sequence of this gene was highly homologous to the coding regions of two rice class I LMM HSP genes, Oshsp 16.9A and Oshsp l6.9B, sharing 93.1% and 94.3% sequence identity, respectively, as published previously in our laboratory. The deduced amino acid sequence of this gene is similar to those of the Oshsp l6.9A and Oshsp l6.9B genes with a difference of only 11 and 10 amino acids, respectively. It was hence designated as Oshsp l6.9C (accession no. U81385). We used 3′ UTRs (untranslated regions) for analysis of Oshsp l6.9C gene expression since the 3′ UTRs of these three genes showed very low sequence homology.

並列關鍵字

Heat shock protein ； Heat shock gene ； Low-molecular-mass heat shock protein ； Oryza sativa L.