| 猪白细胞介素2基因的克隆及腺病毒表达载体的构建 |
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| 引用本文： | 周庆丰,刘忠华,龚朋飞,周美华,苏润环,何津佳,毕英佐.猪白细胞介素2基因的克隆及腺病毒表达载体的构建[J].动物医学进展,2009,30(1). |
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| 作者姓名： | 周庆丰 刘忠华 龚朋飞 周美华 苏润环 何津佳 毕英佐 |
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| 作者单位： | 1. 广东省温氏集团研究院,广东,新兴,527400
2. 华南农业大学动物科学学院,广东,广州,510642 |
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| 摘 要： | 根据GenBank中收录的猪白细胞介素2(pIL-2)基因序列,设计1对特异性引物.运用RT-PCR技术从刀豆素(Con A)诱导的猪外周血淋巴细胞总RNA中扩增得到pIL-2基因,并将其克隆至pGEM-T Easy栽体上.核苷酸测序结果显示,获得的基因序列全长493 bp,含465 bp的开放阅读框,编码154个氨基酸,与已知pIL-2核苷酸序列和氨基酸序列的同源性都高达100%.将克隆的pIL-2基因编码阅读框亚克隆至腺病毒穿梭栽体pShuttle-CMV,电转化含腺病毒基因组(pAdEasy-1)的大肠埃希茵细胞BJ5183-AD-1,成功获得了重组腺病毒DNA,将纯化后的重组腺病毒DNA转染AD-293细胞,经过病毒基因组的PCR鉴定和转录水平的RT-PCR鉴定,初步表明,获得了表达pIL-2的重组腺病毒.
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| 关 键 词： | 猪白细胞介素2 基因克隆 腺病毒载体 基因表达 |
Cloning of Porcine Interleukin-2 Gene and Construction of Recombinant Adenoviral Expression Vector |
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| ZHOU Qing-feng,LIU Zhong-hua,GONG Peng-fei,ZHOU Mei-hua,SU Run-huan,HE Jin-jia,BI Ying-zuo.Cloning of Porcine Interleukin-2 Gene and Construction of Recombinant Adenoviral Expression Vector[J].Progress In Veterinary Medicine,2009,30(1). |
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| Authors: | ZHOU Qing-feng LIU Zhong-hua GONG Peng-fei ZHOU Mei-hua SU Run-huan HE Jin-jia BI Ying-zuo |
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| Institution: | 1.Guangdong Wins Group Academy;Xinxing;Guangdong;527400;China;2.College of Animal Science;South China Agriculture Univversity;Guangzhou;510642;China |
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| Abstract: | Based on the sequences of porcine interleukin 2(pIL-2) gene published in the Genbank,a set of specific primer were designed and synthesized.The cDNA of pIL-2 was amplified by RT-PCR from the total RNA extracted from the peripheral blood lymphocytes which was isolated from Yorkshire sow and stimulated with Concavadine A(Con A),and then the pIL-2 cDNA was cloned into the pGEM-T Easy vector.Sequencing analysis showed that the acquired entire cDNA was 493 bp including the open reading frame(ORF) of pIL-2,which ... |
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| Keywords: | porcine interleukin 2 gene cloning adenoviral vector gene expression |
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