摘要
通过PCR扩增和酶切分别得到抗结肠癌相关抗原抗体的重链可变区序列、轻链可变区序列及连接肽序列,将它们构建成为VHlinkerVL形式的单链抗体基因片段,并在大肠杆菌中进行了表达。SDSPAGE分析结果表明,以pComb3为载体,在大肠肝菌JM83中,scFv未获得有效表达,而以pET22b(+)为载体的scFv在大肠杆菌BL21(DE3)中,30℃诱导培养获得了高效表达,表达水平占全菌蛋白的355%。
Anticolonic cancer scFv fragment with a V H linker V L structure was constructed and expressed in E.coli. SDS-PAGE analysis showed the fragment cloned in pComb3 was not expressed efficiently in JM83, while cloned in pET 22b(+) highly expressed in BL21(DE3)up to 35.5% of the total bacterial protein obtained, if the culture was incubated under 30℃.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第1期82-85,共4页
Chinese Journal of Biotechnology
