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ApiIa抗菌肽基因的化学合成、克隆及其在酵母中的表达 被引量:3
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作者 韩万江 盛泽娟 +3 位作者 张青 宋大新 陈永青 夏天辉 《复旦学报(自然科学版)》 CAS CSCD 北大核心 1998年第2期168-172,共5页
用固相亚磷酰胺法合成了ApiIa抗菌肽基因,全长为80个核苷酸.它被分成4个寡 核苷酸片段,分别在DNA合成仪上合成.经分离纯化后的寡核苷酸片段经酶促连接,然后被 克隆到分泌型载体质粒pFD101上.经限制酶酶切、PC... 用固相亚磷酰胺法合成了ApiIa抗菌肽基因,全长为80个核苷酸.它被分成4个寡 核苷酸片段,分别在DNA合成仪上合成.经分离纯化后的寡核苷酸片段经酶促连接,然后被 克隆到分泌型载体质粒pFD101上.经限制酶酶切、PCR模板检测以及双链 DNA序列分析 检测,证明合成的apiIa基因和设计的完全一致.将含有apiIa基因的质粒pFD1001经BglⅡ 酶切后,转化毕氏酵母.所得转化子经摇瓶发酵,上清液经25%SDS-PAGE检测,有 ApiIa抗 菌肽表达.用抑菌圈试验作定性实验表明,表达的ApiIa有明显的生物活性. 展开更多
关键词 基因 化学合成 ApiIa抗菌肽 无性系 抗菌肽
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STUDIES ON GLUTAMINE SYNTHETASE FROM Streptomyces hygroscopicus var. jinggangensis 被引量:1
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作者 夏天辉 焦瑞身 《Science China Chemistry》 SCIE EI CAS 1986年第4期379-388,共10页
Streptomyces hygroscopicus var. jinggangensis produces validamycin, an important antibiotic used in agriculture. It was found that thexe was a positive correlation between the specific activity of mycelial glutamine s... Streptomyces hygroscopicus var. jinggangensis produces validamycin, an important antibiotic used in agriculture. It was found that thexe was a positive correlation between the specific activity of mycelial glutamine synthetase (GS) and validamycin biosynthesis in our laboratory. So, in this paper, the purification, characteristics and regulatory properties of GS are reported. The native enzyme had a molecular weight of approximately 530,000 and was composed of 12 identical subunits, each 43,000. Electronic microscopic examination of preparations negatively stained disclosed that the subunits were arranged in two hexagonal rings that lay one on top of the other in a face-to-face fashion. Mg2+ or Mn2+ was absolutely needed as cofactor for GS activity. The enzyme activity was regulated by feedback inhibition and cumulative feedback inhibition. In addition, it was also regulated through a covalent modification, adenylylation and deadenylylation, suggesting that the covalent modification of GS exists 展开更多
关键词 jinggangensis STUDIES ON GLUTAMINE SYNTHETASE FROM Streptomyces hygroscopicus var GST
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