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一种高温细菌葡聚糖磷酸化酶基因的克隆与表达 被引量：2

Clone and Overexpression of a Novel Thermostable Glucan Phosphorylase from a Thermophilic Bacterium
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摘要 葡聚糖磷酸化酶(GlgP)在生物体的糖代谢过程中具有中心地位的作用。腾冲嗜热厌氧杆菌(Thermo-anaerobacter tengcongensis MB4T=AS.1.2430T=JCM11007T)分离自云南腾冲的温泉,其最适生长温度为75℃。为研究腾冲嗜热厌氧杆菌的葡聚糖磷酸化酶(Tte-GlgP)的性质,试验以T.tengcongensis MB4T的全基因组为模板,通过PCR扩增得到MB4T中的编码GlgP的基因(glgp),将其克隆到表达载体pET23b上,经过PCR验证、酶切验证和测序验证正确后,转化到宿主细胞大肠杆菌BL21DE3中,成功得到了表达。通过SDS-PAGE电泳分析得到其分子量约为64ku,与预期的结果一致。试验为Tte-GlgP性质的进一步研究和应用构建了基因工程菌株。 Glucan phosphorylase plays a central role in sugar metabolism processing of organism. A novel glucan phosphorylase （Tte-GlgP） from Thermoanaerobacter tengcongensis which was separated from hot spring of Yunnan Tengcong, and its optimum growth temperature is 75℃, was studied in this paper. To characterize the Tte-GlgP, the glgP gene was obtained by polymerase chain reaction （PCR） and was cloned into pET23b. After the colne was validated to be properly constructed by PCR, restrictive enzymes digestions and sequencing methods, it was transformed into E. coli BL21 DE3, and was overexpressed. The high-level expression of Tte-GlgP was confirmed by SDS-PAGE analysis. The Tte-GlgP was a soluble protein, and is about 64 ku as expected.
作者 陈世琼
机构地区 北京市海淀区产品质量监督检验所国家食品质量安全监督检验中心
出处 《食品与发酵工业》 CAS CSCD 北大核心 2009年第7期40-43,共4页 Food and Fermentation Industries
关键词 腾冲嗜热厌氧杆菌 葡聚糖磷酸化酶 克隆 表达 Therrnoanaerobacter tengcongensis, glucan phosphorylase, cloning overexpression
分类号 Q93 [生物学—微生物学] Q78 [生物学—分子生物学]
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参考文献22

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同被引文献18

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食品与发酵工业
2009年 第7期

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