重组人胸腺素β16工程菌株的遗传稳定性研究

目的研究重组人胸腺素β16(Tβ16)工程菌株的遗传稳定性,为重组人Tβ16产业化提供依据。方法利用在有选择压力(AMP+)的条件下,对含有Tβ16-pET表达质粒的工程菌在LB固体斜面上连续传代100代,每隔20代取样进行菌体形态、革兰染色、提取质粒、酶切和进行表达量检测。结果第100代菌株与原始菌株在各项检测指标上一致,无显著差别。结论Tβ16-pET质粒在大肠杆菌BL21(DE3)中有很好的遗传稳定性,为重组人Tβ16的应用奠定了基础。

Study on the genetic stability of recombinant human thymosin β16 bacteria

Purpose In order to verify the genetic stability of recombinant human thymosin β16 bacteria for industrialization of recombinant human thymosin β16.Methods the engineering strain containing Tβ16-pET expression vectors was passaged serially under selection pressure(AMP) on LB agar plates for 100 times,and the tests of bacterial morphology,Gram staining,vector extraction,and the analyses of restriction enzyme and expression were held every 20 generations.Results There was no significant difference between the recombinant bacteria of the 100 generations and the original ones,and the genetic stability of β16-pET expression vector in E.coli BL21(DE3) was quite good.Conclusion It is an important research with significant meanings,which laid a basis for the application of recombinant human thymosin β16 engineering bacteria.