摘要
将含有真养产碱杆菌（Ａｌｃａｌｉｇｅｎｅｓｅｕｔｒｏｐｈｕｓ）合成聚羟基烷酸（ＰＨＡ）基因（ｐｈａＣＡＢ）的质粒ｐＴＺ１８ＵＰＨＢ改造成为具有卡那霉素抗性的质粒ｐＪＭＣ１，并以电击法将ｐＪＭＣ１引入利用碳源广泛的胡罗卜软腐欧文氏菌（Ｅｒｗｉｎｉａｃａｒｏｔｏｖｏｒａ）非致病菌系（Ｅｃｃ１３Ｂ）中，ｐｈａＣＡＢ可获得高效表达，膨大的转基因菌［Ｅｃｃ１３Ｂ（ｐＪＭＣ１）］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。
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
1997年第3期298-303,共6页
Chinese Journal of Biotechnology
基金
国家"八五"重点科技攻关项目
关键词
转基因欧文氏菌
基因表达
聚羟基烷酸
合成酶
Poly β hydroxybutyrate(PHB), phaCAB, transgenic Erwinia , electroporation
