染色体DNA转化原生质球构建解烃工程菌

THE CONSTRUCTION OF ENGINEERING BACTERIA FOR DEGRADATINN HYDROCARBON BY TRANSFORMATING THE SPHEROPLAST WITH CHROMOSOMAL DNA

用供体菌C_(20-11)和D_(36-3)染色体DNA转化受体菌T_3原生质球。采用5mg/ml溶菌酶,37℃水浴1小时制备原生质球。原生质球形成率为99％,再生率为72.5％。在30％、PEG6000和0.1MCaC1_2作用下,供体DNA转化受体原生质球。我们用上述条件构建了能够同时降解以下四种碳源:苯甲酸、萘、樟脑、十六烷的工程菌株TCD32-14-1。

Using the chromosomal DNA of donor strains C20-11 and D36-3 to transform the spheroplast of recipient strain T3, the spheroplast was obtainedby 1hr treatment of lysozyme)5mg (ml) at 37℃。 The rates 1 of fomationand regeneration of the spheroplast were 99％ and 72.5％, respectively. The trensformation was carried out in NSM solution containing 30％ PEG6000, 0.1M CaCl_2. After these treatment, we have constructed the engineering bacteria TCD32-14-1 which can combine the ability to degradate four carbon sources from the parent strains: benzoic acid, naphthalene, hexadecane and camphor. The rate of degradation is also increased.