摘要
目的以人白细胞介素-12(IL-12)双亚基共表达载体pL35P40SN转化大肠杆菌DH5α,并对转化载体进行鉴定。方法以构建好的IL-12双亚基共表达载体pL35P40SN及其对照载体pLXPXSN转化大肠杆菌DH5α,氨苄抗性平板筛选阳性克隆,提取阳性克隆质粒并对其进行酶切及PCR鉴定。结果以XhoI酶切后鉴定,对照载体和IL-12共表达载体的条带分别在6490bp和8800bp的位置,以IL-12 p35和p40引物PCR扩增后电泳鉴定,在700bp和1000bp处可见清亮电泳条带。结论含IL-12 p35和p40双亚基基因的pL35P40SN载体成功转入大肠杆菌DH5α,经多项鉴定结果正确。为以其转染细胞并对IL-12的功能研究打下良好基础。
Objective To transform and identify recombinant human IL-12 co-expression vector pL35P4OSN. Methods Constructed IL-12 co-expression vector was transformed into E. coli DH5α. Extract the plasmid and identify them by digested analysis and PCR. Results Digested with XhoI, there were 6490bp and 8800bp bands that belong to pLXPXSN and pL35P4OSN after agarose gel electrophoresis. IL-12 p35 and p40 primer PCR also showed bands about 700bp and 1000bp. Conclusion The retroviral co-expression vector PL35P4OSN,encoding IL-12 p35 subunit and p40 subunit gene,have transformed into E. coli successfully with identification.
出处
《潍坊医学院学报》
2008年第3期208-210,I0001,共4页
Acta Academiae Medicinae Weifang
基金
山东省卫生厅资助项目(课题编号:HZ154)
