摘要
目的将唾液富组蛋白5 cDNA克隆至表达载体pGEX-4T-1。方法EcoRⅠ+SalⅠ双酶切克隆载体pMD19-T-hrp5及pGEX-4T-1,回收目的片段hrp5及双酶切载体pGEX-4T-1,将二者连接后转化大肠杆菌DH5α,氨苄青霉素筛选阳性菌落,PCR、酶切、测序鉴定重组质粒。结果唾液富组蛋白5的cDNA正确克隆至表达载体pGEX-4T-1,无突变。结论重组载体pGEX-4T-1-hrp5构建成功。
Objective To clone the eDNA of salivary histatin 5 to expression vector pGEX-4T-1. Meth- ods pMD19-T-hrp5 and pGEX-4T-1 were digested by Sal I + EcoR. The digested hrp5 and pGEX-4T-1 were linked and transformed to E. coli DH5α ,and the positive colonies were screened by ampiciUin, and identified by PCR, digestion and sequencing. Results The eDNA of hitatin 5 was correctly inserted into expression vector pGEX-4T-1 without mutation. Conclusion Recombinant vector pGEX-4T-1-hrp5 was constructed successfully.
出处
《咸宁学院学报(医学版)》
2008年第3期191-193,共3页
Journal of Xianning Univarsity(medical Sciences)