【摘要】 目的:通过一系列短引物拼接合成狂犬病毒糖蛋白单链抗体基因Fv57。方法:采用SOE-PCR的方法,利用多条短的寡核苷酸引物,经过6轮PCR,拼接合成800bp左右的狂犬病毒糖蛋白单链抗体基因Fv57,并利用此方法修正了上述PCR过程中产生的多处突变,从而获得正确的单链抗体基因序列。结果:采用SOE-PCR法合成的基因序列经测序及酶切鉴定,与预期结果一致。结论:成功地利用SOE-PCR法合成了狂犬病毒糖蛋白单链抗体基因Fv57,为其下一步构建原核表达载体,在大肠杆菌中进行表达奠定基础。更多还原

【Abstract】 Objective:To synthesize anti-rabies virus G protein scFv fragment gene FV57 through a series of short primer splicing.Method: Utilize the method of SOE-PCR to splice series of short oligonucleotides to synthesize the FV57 gene,and correct the mutations generated from the PCR progress.Result: The results of restriction analysis and genetic sequencing were consistent with those expected.Conclusion: Gene of Fv57 was successfully synthesized by SOE-PCR,which had laid the foundation for subsequent construction of prokaryotic expressing vector,and the expression of anti-rabies virus G protein scFv fragment in E.coli as well.更多还原