摘要
研究了转hTNF-α基因鱼腥藻7120(Anabaenasp.PCC 7120)中表达产物的最佳提取条件,并研究目标蛋白hTNF-α对温度耐受度和硫酸铵盐析的条件,对总蛋白和目标蛋白进行了检测。结果表明,hTNF-α的最佳溶出条件为:0.05 mol/L、pH7.5Tris-HCl缓冲液,NaCl的浓度为0.2 mol/L;当温度为55℃时,处理30min,其hTNF-α提取量下降不明显,回收率达到92.08%,可除去约40%的杂蛋白;先用饱和度为30%的硫酸铵沉淀杂蛋白,再用60%的硫酸铵沉淀目标蛋白,又可除去部分杂蛋白。
This work studied the optimal extracted conditions of hTNF-α from transgenie Anabaena sp. PCC 7120,and the tolerance of hTNF-α to the temperature and the concentrations of ammonium sulphate; detected the concentrations of total proteins and hTNF-α. The results indicated that the optimal extracted conditions. 0. 05mol/L ,pH7. 5Tris-HCl buffer, 0. 2mol/L NaCl; incubated at 55℃, for 30rain,the decrease of the extracted content of hTNF-α was not obvious, and the recovery was about 92.08%,the total proteins of 40% would be removed. 30% ammonium sulphate was used to remove the other protein,60% to deposit hTNF-α,which removed some proteins.
出处
《生物技术通报》
CAS
CSCD
2006年第2期88-92,共5页
Biotechnology Bulletin
基金
天津市科委重点科技攻关项目(043182711)
