摘要
目的：研制ＨＩＶ－１整合酶（ｉｎｔｅｇｒａｓｅ，ＩＮ）的重组噬菌体单链抗体。方法：采用噬菌体表面呈现方法。结果：由ＩＮ蛋白免疫的小鼠脾细胞ｍＲＮＡ中构建了单链抗体基因ｃＤＮＡ文库，克隆入噬菌粒载体ｐＣＡＮＴＡＢ５Ｅ，经转化大肠杆菌ＴＧ１，获得了库容为３．５×１０５的表达文库。利用包被在酶联板上的ＩＮ蛋白进行四轮亲和富集，筛选出７６株具有ＩＮ结合活性的重组噬菌体。对随机挑选的一株克隆进行了ＤＮＡ序列分析，证明其符合小鼠抗体序列。结论：获得了抗ＨＩＶ－１ＩＮ的噬菌体单链抗体，为其进一步研究打下基础。
Objective: Preparation of recombinant phage ScFv against HIV 1 integrase (IN). Methods: The technique of phage surface displaying was used. Results: A combinatory single chain fragment variable (ScFv) cDNA library was constructed from the splenic mRNA of a mouse immunized with IN protein. The library cDNA was cloned into a phagemid vector pCANTAB5E and introduced into E.coli TG1, 3.5×10 5 ampicillin resistant clones were obtained. After four rounds of panning against IN coated on immune plate, 76 IN binding phage ScFvs were selected. DNA Sequence analyzed from a randomly selected clone proved to be in accordance with mouse antibody. Phage ScFvs against HIV 1 IN were collected. Conclusion: This lays built a good foundation for the further study of anti HIV IN ScFvs.
出处
《军事医学科学院院刊》
CSCD
北大核心
1998年第4期248-252,共5页
Bulletin of the Academy of Military Medical Sciences
关键词
艾滋病毒
整合酶
单链抗体
噬菌体
HIV
integrase
ScFv
recombinant phage antibody
bacteriophages
