斜带石斑白细胞cDNA文库的构建

被引：6

作者：

殷志新

翁少萍

叶巧真

何建国

机构：

[1] 中山大学生命科学学院

[2] 中山大学生命科学学院广东广州

[3] 广东广州

来源：

水产学报 | 2001年 / 06期

关键词：

斜带石斑; 白细胞; λTripIEx2; cDNA文库;

D O I：

暂无

中图分类号：

Q785 [转化及克隆];

学科分类号：

071007 ; 0836 ; 090102 ;

摘要：

用Tripure试剂盒提取斜带石斑白细胞总RNA ,反转录合成第一链cDNA ,进行长距离PCR ,蛋白酶K消化 ,SfiI酶切 ,通过CHROMASPIN - 4 0 0柱 ,回收大于 5 0 0bp的cDNA ,与λTripIEx2载体连接 ,体外包装后构建了斜带石斑白细胞的cDNA文库。库容 1.6 5× 10 6,重组频率 82 % ,扩增后滴度 7.5× 10 9pfu·mL-1。插入片断长度 5 0 0～ 2 30 0bp ,最多的是在 75 0～ 10 0 0bp范围。本文库可作为筛选斜带石斑细胞因子基因的重要资源

引用

页码：538 / 541

页数：4

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