产谷氨酸棒杆菌B9和T6—13的原生质体融合

Protoplast Fusion Of Corynebacterium B9 and T6 - 13 Strains of Producing Glutamic Acid

棒杆菌Ｂ９和Ｔ６—１３两菌株经ＵＶ变处理得到Ｂ９—２（ＳｍＲ）和Ｔ６—１３—３（ＲｉｆＲ）两菌株，以此两菌株做为出发菌株。将对数前期的培养细胞经青霉素予处理及酶解制备原生质体，用４０％ＰＥＧ６０００为助融剂，进行原生质体融合。用间接法检出具有Ｓｍ、Ｒｉｆ抗性的融合子。融合频率为６．５５×１０－６－１．６４×１０－５，融合子双抗性稳定，产谷氨酸，经摇瓶实验筛选出一株产谷氨酸明显高于亲本的融合子ｆｕ３６。

Two mutant strains, B2(Smr)and T3(Rifr),Were obtained by UV mutagenesis of corynebacterinm b9 and t6-13.USing them as parent strains, the cultural Cells in early togarithmic phase was treated by penicillin firstly and then lysozyme, The protoplast were prepared.Protoplast fusion was made with the help of 40 % PEG6000, the fusants that had the resistance to sm and Rif were detectedby indirect method. The frequency of fusion was 6. 55 × 10-6-1. 64 × 10 × -5. Double resistance of the fusants is stable. By shake flat experiment, fusant fu36 was selected and amount of glutamic acid that produced were obriously higher than Parent strains.