关键词: 胸腺素α1 ;串联体;克隆;基因表达;融合蛋白

    摘 要:目的 利用基因工程方法表达胸腺素α1 (thymosin alpha1，Tα1 )的串联体并进行纯化、鉴定. 方法 将Tα1 的基因拆分为4个片段进行合成，退火后经PCR获得串联体基因，测序正确后克隆在硫氧还蛋白(thioredoxin)融合蛋白表达载体pThioHisA中，转化大肠杆菌TOP10菌株，IPTG诱导表达，表达的融合蛋白经过80℃热处理及阴离子交换柱Q-Sepharose Fast Flow进行纯化，Western-blot进行鉴定. 结果 获得了纯化的Thioredoxin-Tα1 ③，分子质量约为31ku. 结论 用基因串联思路表达小分子多肽是一种可行的方法，Tα1 ③的成功表达为检测其生物学活性奠定了基础.

    Cloning，expression and purification of recombi┐nant thymosin alpha1

    XUE Xiao-Chang，YAN Zhen，SHI Ji-Hong，HAN Wei，ZHANG Ying-Qi

    Biotechnology Centre，Fourth Military Medical U-niversity，Xi'an710033，China

    Keywords:thymosinα1 ;tandem repeat;cloning;gene ex-pression;fusion proteins

    Abstract:AIM To express tandem repeat of thymosin alpha1(Tα1 )with gene engineering method and to purify and iden-tify the fusion protein.METHODS Split Tα1 gene into four fragments，synthesize them respectively and get the tandem repeat gene of Tα1 by annealing and PCR.After sequencing to identify its correctness，clone the gene into fusion expres-sion vector pThioHisA，transforms the E.coli strain TOP10with recombinant plasmid and induces with IPTG.The ex-pressed fusion protein can be purified by heat treatment and Q Sepharose Fast Flow ion exchange chromatography and i-dentified by western-blot.RESULTS We obtained purifiedThioredoxin-Tα1 ③the molecular weight of which was about31ku.CONCLUSION To express short peptide ......