【摘要】 应用RT-PCR技术克隆犬IL-2基因,并插入到原核表达载体P JLA 605中,构建pRL-C aIL-2表达质粒;采用M 9培养基摇瓶发酵,确定诱导时机和诱导表达时间。结果表明:工程菌pRL-C aIL-2在30℃培养至对数生长后期(OD600为1.5)42℃诱导4 h时,菌体收得量湿重达17.8 g/L,目标蛋白表达量约占菌体总蛋白的29.7%。外源基因在该基因工程菌中得到了高效表达。更多还原

【Abstract】 The canine IL-2 gene of was cloned by RT-PCR and was inserted into PJLA605 vector.The recombinant plasmid of pRL-CaIL-2 was constructed.The cell growth phases for induction and inducing period were determined using shaking flask with M9 medium.The results showed that when induced at 42℃ for 4 hours after culture the E.coli DH_5α（pRL-CaIFN-γ） in shaking flask reached to 1.5(OD₆₀₀) at 30℃,17.8 g of wet bacteria per liter could be obtained and the derivative of CaIL-2 was about 29.7% of total protein in the host.The gene was highly expressed in the engineering bacterial strain.更多还原