薯蓣皂苷糖苷酶基因的克隆及同源性分析 |
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引用本文: | 吴宁,金凤燮,鱼红闪.薯蓣皂苷糖苷酶基因的克隆及同源性分析[J].安徽农业科学,2011,39(8):4438-4439,4442. |
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作者姓名: | 吴宁 金凤燮 鱼红闪 |
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作者单位: | 大连工业大学生物工程学院,辽宁大连,116034;大连工业大学生物工程学院,辽宁大连,116034;大连工业大学生物工程学院,辽宁大连,116034 |
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基金项目: | 国家自然科学基金项目(30371744,30470055,20476017);辽宁省科学技术基金项目(20042132);辽宁省教育厅创新团队项目(2009T007) |
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摘 要: | 目的]从分子生物学水平上研究Absidia sp.G3d产薯蓣皂苷糖苷酶,对该酶基因CDS区的序列进行调取和分析。方法]根据薯蓣皂苷糖苷酶的N端序列设计简并引物,以总RNA为模板进行RT-PCR调取cDNA,然后采用3'RACE和5'RACE技术,调取CDS区基因全序列。结果]测序得到序列长度为1 497 bp。该基因的同源性比较分析表明,薯蓣皂苷糖苷酶基因与α-鼠李糖苷酶没有同源性,与α-淀粉酶的基因序列具有很高的同源性,达到93%以上。结论]蛋白质结构存在差异,酶反应特性不同。
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关 键 词: | 薯蓣皂苷糖苷酶 RT-PCR 序列分析 |
Cloning and Homology Analysis of Dioscin-glycosidase Gene from Absidia sp.G3d |
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Institution: | WU Ning et al(College of Bioengineering,Dalian Polytechnic University,Dalian,Liaoning 116034) |
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Abstract: | Objective] A dioscin-glycosidase from Absidia sp.G3d was studied in molecular biology.Dioscin-glycosidase gene of CDS was cloned and analyzed.Method] The primers were designed according to N-terminal sequence of dioscin-glycosidase.The partial cDNA of dioscin-glycosidase gene was obtained through the method of RT-PCR using total RNA as template.Moreover,a 1 497 bp full length CDS sequence of dioscin-glycosidase was obtained using RT-PCR,5'-RACE and 3'-RACE technique.Sequence homology analysis of dioscin-glycosidase gene was performed by BLAST and subsequently compared with GenBank data.Result] There was no identity between dioscin-glycosidase gene and the known α-rhamnosidase gene in the Genbank.But there was a high homogeneity with alpha-amylase.Conclusion] There was difference between the protein structures.And the reaction characteristics of the protein were different. |
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Keywords: | Dioscin-glycosidase RT-PCR Sequence analysis |
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