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利用SMART技术构建鸡法氏囊全长cDNA表达文库 被引量：2

Full length cDNA expression library construction of bursa of fabricius in chicken by SMART technology
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摘要 为构建鸡法氏囊组织互补DNA(cDNA)文库,从3周龄鸡法氏囊组织提取总RNA,用偶联oligo(dT)磁珠纯化mRNA,用SMART(Switching methanism at 5′end of RNA transcrip)技术合成cDNA第1链,用LD-PCR方法扩增双链cDNA。经SfiⅠ酶切消化后,用CHROMA SPIN-400柱去除400 bp以下小片段,获得的双链cDNA与同样酶切的pMyr表达载体连接,电转化大肠杆菌获得cDNA文库。初始文库容量为2.8×10~6cfu,重组子比例为100%,插入片段的大小平均为1.2 kb。扩增后文库容量为8×10^(11)cfu·mL^(-1)。所建文库为鸡传染性法氏囊病毒受体基因克隆及功能研究奠定基础。 In order to clone and characterize IBDV receptor gene(s),mRNAs were isolated from the bursa of Fabricius of 3 week-old SPF chicken for construction of cDNA library using SMART technique.The first strand cDNAs were synthesized by reverse transcription using the CDSⅢprimer with Sfi I site introduced and the double-stranded cDNAs were amplified by Long-distance-PCR.After removing small fragments less than 400 bp,the cDNAs were ligated with the pMyr vector in the CytoTrap^(TM) Two-Hybrid System and transformed ...
作者 袁维峰 吴保明 张鑫宇 张泉 孙怀昌
机构地区 扬州大学兽医学院
出处 《扬州大学学报（农业与生命科学版）》 CAS CSCD 北大核心 2009年第1期1-4,共4页 Journal of Yangzhou University：Agricultural and Life Science Edition
基金 国家自然科学基金资助项目(30571373)
关键词 鸡 法氏囊组织 CDNA表达文库 鸡传染性法氏囊病毒 chicken the bursa of Fabricius cDNA expression library IBDV
分类号 S858.31 [农业科学—临床兽医学]
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共引文献29

同被引文献33

  • 1崔言顺,李建亮,刘星火,姜世金,孙淑红,崔治中,张彦明.传染性法氏囊病超强毒山东分离株SD0210的致弱研究[J].中国病毒学,2005,20(4):408-412. 被引量：3
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扬州大学学报（农业与生命科学版）
2009年 第1期

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