cDNA cloning and expression pattern of two enolase genes from the Chinese oak silkworm, Antheraea pernyi
cDNA cloning and expression pattern of two enolase genes from the Chinese oak silkworm, Antheraea pernyi
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柞蚕两个烯醇化酶基因的cDNA克隆及表达模式
DOI：
10.1093/abbs/gmq084
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发表时间：
2010-11-01
影响因子：
3.7
通讯作者：
Xiang, Zhonghuai
中科院分区：
文献类型：
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作者：
Liu, Yanqun;Li, Yuping;Xiang, Zhonghuai
本研究从中国柞蚕中分离并鉴定了两个烯醇化酶基因，分别命名为烯醇化酶I和烯醇化酶II。该基因cDNA全长1712 bp，开放阅读框（ORF）为1302 bp，编码433个氨基酸。烯醇化酶Ⅱ cDNA序列全长1549 bp，开放阅读框1296 bp，编码431个氨基酸。这两个基因的氨基酸序列共享烯醇化酶的几个保守特征/位点。柞蚕烯醇化酶I与鳞翅目昆虫烯醇化酶的同源性为93%-97%，与其它无脊椎动物烯醇化酶的同源性为75%-82%，与其它生物包括脊椎动物、植物和真菌的烯醇化酶的同源性为60%-72%。柞蚕烯醇化酶II与家蚕烯醇化酶II的同源性为84%，而与家蚕烯醇化酶II的同源性为60%。紫苏烯醇化酶I在系统进化树中，A. pernyi和B. mori的烯醇化酶序列与包括A. pernyi和B.桑属烯醇化酶I序列。通过序列比较和系统进化分析，我们认为A. pernyi和B. Mori可能是烯醇化酶超家族的新成员。柞蚕烯醇化酶I基因在所有组织中均有表达，烯醇化酶II基因在精巢和卵巢中特异表达，表明烯醇化酶II基因产物可能与生殖有关。A.冷激后，紫苏烯醇化酶I基因表达显著下调，热激后，紫苏烯醇化酶I基因表达显著上调，提示紫苏烯醇化酶I基因可能与冷激有关。紫苏烯醇化酶I基因可能受温度胁迫诱导。
In this study, two enolase genes were isolated and characterized from the Chinese oak silkworm, Antheraea perny, which were designated as enolase I and II, respectively. The enolase I cDNA sequence was 1712 bp with an open reading frame (ORF) of 1302 bp encoding 433 amino acids. The enolase II cDNA sequence was 1549 bp with an ORF of 1296 bp encoding 431 amino acids. The amino acid sequences of the two genes share several conserved features/sites of enolase. Antheraea pernyi enolase I shows 93%-97% sequence identity to enolases of lepidopterans available to date, 75%-82% identity to enolases of other invertebrates, 60%-72% identity to enolases of other organisms including vertebrates, plants, and fungi. Antheraea pernyi enolase II shows 84% identity to Bombyx mori enolase II, but 60% identity to A. pernyi enolase I. In the phylogenetic tree, enolase II sequences from A. pernyi and B. mori were clearly separated from the majority of enolase sequences of higher organisms including A. pernyi and B. mori enolase I sequences. By sequence comparisons and phylogenetic analysis, we suggest that enolase II from A. pernyi and B. mori may be a new member of the enolase superfamily. Antheraea pernyi enolase I mRNA was found in all tested tissues whereas enolase II mRNA was expressed specifically in the spermaries and ovaries, suggesting that the product of enolase II gene may be related to reproduction. The transcript abundance of A. pernyi enolase I gene was significantly down-regulated after cold shock and significantly up-regulated after heat shock, suggesting that A. pernyi enolase I gene may be inducible by temperature stress.