| 利用iCODEHOP设计简并引物克隆益生菌FGM通透酶基因片段 |
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| 引用本文： | 王龙,张凯,王旭荣,张景艳,孟嘉仁,郝桂娟,杨志强,李建喜.利用iCODEHOP设计简并引物克隆益生菌FGM通透酶基因片段[J].中国畜牧兽医,2012,39(8):64-67. |
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| 作者姓名： | 王龙 张凯 王旭荣 张景艳 孟嘉仁 郝桂娟 杨志强 李建喜 |
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| 作者单位： | 中国农业科学院兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,甘肃兰州 730050 |
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| 基金项目： | 国家自然科学基金项目，中央级公益性科研院所基本科研业务费专项资金项目 |
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| 摘 要： | 作者通过iCODEHOP在线设计细菌通透酶的简并引物,以发酵黄芪菌FGM基因组DNA为模板进行TouchdownPCR扩增,得到740 bp PCR产物,将产物经pGEM-T Easy载体连接,转化至JM109中,筛选阳性株并测序。序列通过BLAST x检索与GenBank进行同源性比对后,结果表明,此DNA产物序列与其他菌属来源的通透酶蛋白序列具有相似性,所克隆的序列即为FGM通透酶基因片段。用iCODEHOP在线设计的简并引物可信性强,阳性率高。FGM通透酶基因的成功克隆为细菌发酵黄芪机理研究提供了依据。
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| 关 键 词： | 益生菌 iCODEHOP 通透酶 |
The Cloning of Permease Gene Fragment from Probiotics FGM Using Degenerate Primers Designed with iCODEHOP |
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| WANG Long
,
ZHANG Kai
,
WANG Xu-rong
,
ZHANG Jing-yan
,
MENG Jia-ren
,
HAO Gui-juan
,
YANG Zhi-qiang
,
LI Jian-xi.The Cloning of Permease Gene Fragment from Probiotics FGM Using Degenerate Primers Designed with iCODEHOP[J].China Animal Husbandry & Veterinary Medicine,2012,39(8):64-67. |
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| Authors: | WANG Long ZHANG Kai WANG Xu-rong ZHANG Jing-yan MENG Jia-ren HAO Gui-juan YANG Zhi-qiang LI Jian-xi |
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| Institution: | (Engineering and Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province,Lanzhou Institute of Animal Science and Veterinary Pharmaceutics,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China) |
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| Abstract: | Degenerate PCR for permease using iCODEHOP to design the degenerate primers,chosing a pair of degenerate primers and utilizing the probiotics bacteria genome DNA as template.740 bp PCR product was obtained,transformed into the E.coli JM109 through being linked with pGEM-T easy vector and sequenced after filtration.Similarity alignment showed that the products of the cloned DNA were similar to those of permease gene from the other strains.The cloned sequence was putatively permease gene DNA fragment from FGM strain.The results indicated that the degenerate primers designed by the iCODEHOP software could be used to obtain specific gene fragment.Cloning of gene fragment would give a scientific warrant for further study of the fermenting mechanism of astragalus. |
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| Keywords: | probiotics iCODEHOP permease |
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