| 真养产碱杆菌聚羟基烷酸合成酶基因在欧文氏菌中的表达 |
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| 引用本文： | 田杰生,李季伦.真养产碱杆菌聚羟基烷酸合成酶基因在欧文氏菌中的表达[J].生物工程学报,1997,13(3):298-303. |
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| 作者姓名： | 田杰生 李季伦 |
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| 作者单位： | 中国农业大学生物学院微生物系 北京 100094 |
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| 基金项目： | 国家“八五”重点科技攻关项目 |
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| 摘 要： | 将含有真养产碱杆菌(Alcaligenes eutrophus)合成聚羟基烷酸(PHA)基因(phaCA3)的质粒pTZl8U—PHB改造成为具有卡那霉素抗性的质粒pJMCl．并以电击法将pJMCl引入利用碳源广泛的胡罗卜软腐欧文氏茵(Erwinia carotovora)非致病菌系(Eccl3B)中，phaCAB可获得高效表达，膨大的转基因菌Eccl3B(pJMCl)]细胞内几乎充满PHA颗粒。以蔗糖为碳源，初步在5L发酵罐中对转基因菌进行分批补料培养35h，菌体干重达28g／L,PHA占菌体干重的68％，具有生产潜力。将该菌合成的PHA提取纯化(纯度达99％)后，进行核磁共振分析，发现它只有单一的聚-β-羟基丁酸(PHB)组分。
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| 关 键 词： | 聚-β-羟基丁酸，PHA合成酶基因．转基因欧文氏菌，电击转化 |
Expression of phaCAB Genes in Transgenic Erwinia carotovora 13B |
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| Tian Jiesheng\ Li Jilun.Expression of phaCAB Genes in Transgenic Erwinia carotovora 13B[J].Chinese Journal of Biotechnology,1997,13(3):298-303. |
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| Authors: | Tian Jiesheng\ Li Jilun |
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| Abstract: | A derived plasmid, pJMC1, containing ployhydroxyalkanote (PHA) synthetic genes (phaCAB) from Alcaligenes eutrophus was successfully introduced into a non pathogenic strain of Erwinia carotovora (Ecc13B) by electroporaiton. The pahCAB genes were expressed well in the transgenic bacteria, Ecc13B (pJMC1). Fed batch cultures of the transgenic Erwinia were conducted in a 5L automatic control fermentor with sucrose as the carbon source, and it shew that the final cell mass and PHB content of dry cell weight were 28g/L and 68% respectively within 35 hours. |
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