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| SOE-PCR法合成狂犬病毒单链抗体基因Fv57 | |
| 引用本文： | 谷铁军,张凤羽,段冶,卫巍,姜春来,吴永革,孔维.SOE-PCR法合成狂犬病毒单链抗体基因Fv57[J].生物技术,2011,21(3). |
| 作者姓名： | 谷铁军 张凤羽 段冶 卫巍 姜春来 吴永革 孔维 |
| 作者单位： | 1. 吉林大学生命科学学院艾滋病疫苗国家工程实验室,吉林长春,130012 2. 长春百克生物科技股份公司,吉林长春,130012 3. 吉林大学生命科学学院艾滋病疫苗国家工程实验室,吉林长春,130012;长春百克生物科技股份公司,吉林长春,130012 |
| 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. |
| 关 键 词： | 单链抗体基因 基因突变 |
Synthesis of Anti-Rabies ScFv Fragment Gene FV 57 by SOE-PCR |
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| GU Tie-jun,ZHANG Feng-yu,DUAN Ye,WEI Wei,JIANG Chun-lai,WU Yong-ge,KONG Wei.Synthesis of Anti-Rabies ScFv Fragment Gene FV 57 by SOE-PCR[J].Biotechnology,2011,21(3). | |
| Authors: | GU Tie-jun ZHANG Feng-yu DUAN Ye WEI Wei JIANG Chun-lai WU Yong-ge KONG Wei |
| Institution: | GU Tie-jun1,ZHANG Feng-yu2,DUAN Ye1,WEI Wei1,JIANG Chun-lai1,2,WU Yong-ge1*,KONG Wei1*(1.National Engineering Laboratory for AIDS Vaccine,College of Life Science,Jilin University,Changchun 130012,China,2.Changchun BCHT Biotechnology Co.,China) |
| 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 o... |
| Keywords: | SOE-PCR |
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