【摘要】：產(chǎn)黃青霉(Penicillium chrysogenum)是?-內(nèi)酰胺類抗生素的主要工業(yè)生產(chǎn)菌,高效地對其基因進行改造有利于深入研究產(chǎn)黃青霉的代謝機制,進而提高效價、產(chǎn)率和得率。高產(chǎn)產(chǎn)黃青霉基因測序的完成為菌株定向改造提供了分子基礎,但是要實現(xiàn)快速的定向菌株改造還需要高效的基因操作系統(tǒng)。本試驗采用土壤農(nóng)桿菌(Agrobacterium)轉化法高效構建了海藻糖循環(huán)途徑tps1、tps2敲除菌株P.chrysogenum-Δtps1和P.chrysogenum-Δtps2。土壤農(nóng)桿菌為高毒力的根癌農(nóng)桿菌AGL-1,載體為p Green II-0179,篩選標記為博來霉素,轉化對象為產(chǎn)黃青霉孢子。在本試驗條件下農(nóng)桿菌轉化法的轉化效率可達60%,其中發(fā)生同源重組的效率為25%。本研究為后續(xù)探究海藻糖循環(huán)在高產(chǎn)產(chǎn)黃青霉中的生理學意義提供了有益的參考。
[Abstract]:Penicillium luteum (Penicillium chrysogenum) is the main industrial producing strain of lactam antibiotics. It is helpful to study the metabolic mechanism of Penicillium xanthogenes and to improve its titer, yield and yield by modifying its gene efficiently. The completion of gene sequencing of high-yielding Penicillium flavus provides a molecular basis for the directed transformation of the strain, but the efficient gene operating system is needed to realize the rapid transformation of the directed strain. The trehalose cycle pathway tps1,tps2 knockout strains P.chrysogenum- 螖 tps1 and P.chrysogenum- 螖 tps2. were efficiently constructed by Agrobacterium tumefaciens (Agrobacterium) transformation in this experiment. Agrobacterium tumefaciens was a highly virulent Agrobacterium tumefaciens AGL-1, vector with p Green II-0179, screening marker as bleomycin and transformed to Penicillium flavum spores. In this experiment, the transformation efficiency of Agrobacterium tumefaciens transformation method can reach 60%, and the efficiency of homologous recombination is 25%. This study provides a useful reference for further exploring the physiological significance of trehalose cycle in high yield penicillium luteum.
【作者單位】： 華東理工大學生物反應器國家重點實驗室;
【基金】：中荷國際合作項目(No.2013DFG32630)給予資助
【分類號】：R915

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