| 总状毛霉（Mucor racemosus）甲壳素脱乙酰酶全长cDNA的克隆及其系统发生分析 |
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| 引用本文： | 蒋霞云,邹曙明,周培根.总状毛霉（Mucor racemosus）甲壳素脱乙酰酶全长cDNA的克隆及其系统发生分析[J].农业生物技术学报,2006,14(6). |
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| 作者姓名： | 蒋霞云 邹曙明 周培根 |
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| 作者单位： | 上海水产大学食品学院 上海水产大学农业部水产种质资源与养殖生态重点开放实验室 |
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| 摘 要： | 用保守的特异引物进行PCR扩增，并结合RACE技术，分离到了总状毛霉（Mucor racemosus）甲壳素脱乙酰酶(CDA)基因，并进行了全序列测定，并提交GeneBank（DQ538514）。研究结果表明：（1）总状毛霉CDA基因全长为1506 bp，包括67 bp 5’非翻译区，1357 bp阅读框以及82bp 3’非翻译区，3’非翻译区包含Poly (A) 加尾信号AATAAA。总状毛霉的CDA基因共编码448个氨基酸，在该基因中部还包含一个144氨基酸的多糖脱乙酰酶结构域，约占CDA基因全长的32%。（2）总状毛霉CDA基因与其它相近种米根霉（Rhizopus oryzae）、卷柄根霉（Rhizopus circinans）的CDA1和CDA2、鲁氏毛霉（Mucor rouxii）、卵形孢球托霉（Gongronella butleri）、匍枝根霉（Rhizopus stolonifer）、布拉克须霉（Phycomyces blakesleeanus）、酿酒酵母（Saccharomyces cerevisiae）的CDA1和CDA2的基因序列同源性分别为：75%、58%、56%、56%、48%、39%、39%、17%和16%；相应的氨基酸序列的同源性分别为：69.0%、57%、59%、55%、47%、30%、32%、18%、21%。表明CDA基因在不同的真菌中有较高的同源性。（3）根据总状毛霉CDA基因的氨基酸序列，构建的不同真菌的系统树，与采用经典分类法构建的系统树基本一致。（4）通过生物信息学的方法，预测该基因所编码的蛋白质三级结构，验证了该蛋白质具有甲壳素脱乙酰酶完整的功能性结构，并包含一个含多糖脱乙酰酶结构域，两者具有相似的空间结构。
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| 关 键 词： | 快速扩增cDNA末端 甲壳素脱乙酰酶 总状毛霉 系统发生 |
| 收稿时间： | 2006-6-23 |
| 修稿时间： | 2006-9-12 |
Cloning complete cDNA of chitin deacetylase from Mucor racemosus and its phylogenetic analysis |
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| Abstract: | A complete chitin deacetylase (CDA) cDNA from Mucor racemonus was cloned and sequenced by using RT-PCR and RACE with special conserved primers. The cDNA sequence was submitted to GeneBank (DQ538514). The results showed: (1) The complete gene with length of 1506bp contains 67bp 5’-untranslated region, an open reading frame of 1357bp and 82bp 3’-untranslated region including tailing site AATAAA. The gene encodes a sequence of 448 amino acid residues and consists of core nucleotides encoding a polysaccharide deacetylase domain which covering 32 of the entire sequence. (2) The CDA gene shares high sequence similarity with that of fungi including Rhizopus oryzae (75%), CDA1(58%) and CDA2(56%) of Rhizopus circinans, Mucor rouxii(56%), Gongreonella bulteri(48%), Rhizopus stolonifer(39%), Phycomyces blakesleeanus(39%), CDA1(17%) and CDA2(16%) of Saccharomyces cerevisiae. The corresponding homology of the deduced amino acid sequences was 69%, 57%, 59%, 55%, 47%, 30%, 32%, 18% and 21%, respectively. (3) Phylogenetic analysis according to the deduced amino acid sequences was marched with the classical taxonomic classification of the fungi. (4) The 3-D-structure of this protein was predicted. The protein has a whole CDA functional domain and a polysaccharide deacetylase domain. |
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