文章摘要
殷志新.斜带石斑白细胞cDNA文库的构建[J].水产学报,2001,25(6):
斜带石斑白细胞cDNA文库的构建
Construction of cDNA library of Epinephelus coioides leukocytes
投稿时间:2014-04-14  修订日期:2014-04-14
DOI:
中文关键词: 斜带石斑 白细胞 cDNA文库 石斑鱼
英文关键词: Epinephelus coioides  Leucocytes,KTripIEx 2 , cDNA Library
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作者单位地址
殷志新 中山大学生命科学学院 中山大学生命科学学院
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中文摘要:
      斜带石斑 白细胞 cDNA文库 石斑鱼
英文摘要:
      Total RNA of leukocytes from orange-spotted grouper, Epinephelus coioides were extracted by Tripure kit. The first strand cDNA was synthesized by using MMLV reverse transcriptase. Then the cDNA were amplif ied by LD- PCR with SMART ó Oligonucleotide and CDS ó as Primers. After proteinase K digestion , Sfi . digestion, chromatography through CHROMASPIN- 400, over 500bp fragments were collected and ligated with KTripIEx2 vector. After package in vitro, cDNA library containing 1. 65 @ 106 private clones was constructed, in which more than 82% clones were recombinant . After amplif ication, the titer of the library was 7. 5 @ 109 pfu#mL- 1. The insert DNA were between 500 and 2300bp, and most of them were between 750 and 1000bp. The library can be used to screen cytokine genes of orange-spotted grouper.
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