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当前位置: 首页 > 文章 > 抗鹅α-干扰素单链抗体基因扩增与表达 河南农业科学 2016,45 (8) 136-139
Position: Home > Articles > Amplification and Expression of the Single-chain Fv Gene Specific for Goose IFN-α Journal of Henan Agricultural Sciences 2016,45 (8) 136-139

抗鹅α-干扰素单链抗体基因扩增与表达

作  者：
姚志兰;邓祖丽颖;傅宏庆;黄平;江明香;刘永鑫;王永娟
单  位：
阜宁县鑫永生态饲养场;江苏农牧科技职业学院江苏省兽用生物制药高技术研究重点实验室;郑州幼儿师范学校;安吉以琳生物科技有限公司;湖州市安吉县畜牧兽医局
关键词：
鹅;α-干扰素;单链抗体;测序;表达
摘  要：
为制备抗鹅α-干扰素的单链抗体,以抗鹅α-干扰素的杂交瘤细胞株总RNA为模板,RT-PCR法扩增鹅α-干扰素的抗体轻、重链基因,再采用SOE-PCR法,以编码柔性多肽(Gly4Ser)3为接头,组装出完整的鹅α-干扰素的单链抗体可变区片段(Sc Fv)基因,并将Sc Fv基因克隆到p GEMT-Easy载体中进行测序分析,将测序正确的Sc Fv基因片段克隆入p ET-30a载体中进行诱导表达。结果显示,成功组装了Sc Fv基因,其全长为726 bp,为VH-Linker-VL结构,其中VH长357 bp、VL长324 bp,成功表达了Sc Fv蛋白,大小为27 ku。
译  名：
Amplification and Expression of the Single-chain Fv Gene Specific for Goose IFN-α
作  者：
YAO Zhilan;DENGZU Liying;FU Honqing;HUANG Ping;JIANG Mingxiang;LIU Yongxin;WANG Yongjuan;Jiangsu Agri-animal Husbandry Vocational College,Jiangsu Provincial Key Laboratory of Veterinary Biopharmaceutical High-tech Research;Zhengzhou Infant Normal School;Animal Husbandry and Veterinary Bureau of Anji;Anji to Lin Biological Technology Co.Ltd.;Funing County Xinyong Ecological Farms;
关键词：
goose;;IFN-α;;single chain variable fragment;;sequencing;;expression
摘  要：
In order to obtain the single chain variable fragment( Sc Fv) gene specific for goose IFN-α,total light and heavy domains of the immunoglobulin were amplified from the RNA isolated from anti goose IFN-α monoclonal hybridoma cells by RT-PCR method. Full length of Sc Fv was assembled by overlap extension PCR with a linker containing flexible polypeptide( Gly4Ser)3. The Sc Fv fragment was cloned into the vector p GEMT-Easy and sequenced. And then it was cloned into p ET-30 a for expression. As a result,the Sc Fv gene related to goose IFN-α,consisting of 726 bp with the structure of VH-Linker-VL was successfully amplified. The VH gene was 357 bp,while the VL gene was 324 bp. Sc Fv could be expressed in p ET-30a(27 ku).

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