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| Geobacillus sp.POT5 α-淀粉酶基因的克隆及表达 | |
| 引用本文： | 杨灵,侯瑛,余霖霖,刘国生,张志芳.Geobacillus sp.POT5 α-淀粉酶基因的克隆及表达[J].河南师范大学学报(自然科学版),2007,35(3):135-139. |
| 作者姓名： | 杨灵 侯瑛 余霖霖 刘国生 张志芳 |
| 作者单位： | 1. 河南师范大学,生命科学学院,河南,新乡,453007;中国农业科学院,生物技术研究所,北京100081 2. 河南师范大学,生命科学学院,河南,新乡,453007 3. 中国农业科学院,生物技术研究所,北京,100081 |
| 摘 要： | 从高温环境土壤中分离到1株能在75℃生长并产生α-淀粉酶的菌株(POT5),扩增了其16Sr DNA核苷酸序列,序列分析表明该菌属于Geobacillus属.根据该属全基因序列测定数据中推定的α-淀粉酶基因,利用PCR方法从G.sp.POT5基因组中扩增得到该菌α-淀粉酶基因(amyP).序列分析表明该基因全长1.545 kb、G+C含量51.33%,编码514个氨基酸.构建重组表达质粒pET22b(+)-amyP,转化Escherichia coliBL21系统,表达产物经SDS-PAGE分析、活性染色及淀粉酶活力分析,表明amyP基因得到了表达,且产物具有生物学活性. |
| 关 键 词： | Geobacillus 16SrDNA 淀粉酶 基因表达 |
| 文章编号： | 1000-2367（2007）03-0135-05 |
| 修稿时间： | 2007-01-16 |
Cloning, Sequencing, and Expression of α-amylases Gene from Geobacillus sp. POT5 |
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| YANG Ling,HOU Ying,YU Lin-lin,LIU Guo-sheng,ZHANG Zhi-fang.Cloning, Sequencing, and Expression of α-amylases Gene from Geobacillus sp. POT5[J].Journal of Henan Normal University(Natural Science),2007,35(3):135-139. | |
| Authors: | YANG Ling HOU Ying YU Lin-lin LIU Guo-sheng ZHANG Zhi-fang |
| Institution: | 1. College of Life Sciences, Henan Normal University, Xinxiang 453007,China;2, Biotechnology Research Institute, Chinese Academy of Agricultural Science, Beijing 100081,China |
| Abstract: | A strain of bacterium which can grow at the temperature of 75 ℃ and secrete α-amylases was isolated from soil of thermal environment.It was identified as Geobacillus sp.according to the analysis of 16S rDNA nucleotide sequence.The gene of α-amylases(amyP)was amplified from the genome of G.sp.POT5 by PCR.Sequence analysis indicated that the gene was 1 545 bp in full size with 51.33% content of G+C,encoding 514 amino acids.The recombinant expression vector pET22b(+)-amyP was constructed and transformed into Escherichia coli BL21,the expressed product was detected through SDS-PAGE,active-staining and activity analysis of α-amylase. |
| Keywords: | Geobacillus 16Sr DNA u-amylases gene expression |
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