SOE-PCR法合成狂犬病毒单链抗体基因Fv57

Synthesis of Anti-Rabies ScFv Fragment Gene FV 57 by SOE-PCR

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。

Objective：To synthesize anti -rabies virus G protein scFv fragment gene FV57 through a series of short primer splicing. Meth- od： 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 pro- karyotic expressing vector, and the expression of anti - rabies virus G protein scFv fragment in E. coli as well.