大麻哈鱼基因文库的构建及其生长激素基因的克隆  

Study on Construction of Salmon Genomic DNA Library and the Cloning of Salmon Growth Hormone Gene

作　　者：杨学成[1] 杨志兴[1] 刘伟民[1] 毕吉威 赵晓祥[1] 姜凯波[1] 张云湖[1] 李洪超[1] 

机构地区：[1]黑龙江省科学院应用微生物研究所 [2]山东省威海市医药公司

出　　处：《生物技术》1991年第1期13-16,共4页Biotechnology

基　　金：黑龙江省科学院资助

摘　　要：本文主要论述了黑龙江省特有鱼种,大麻哈鱼全基因文库的构建方法以及从所构建的1.9×10^(?)噬菌体成斑单位/ugDNA基因文库中克隆出生长激素基因片段的研究结果,作者使用DM-BL3入噬菌为载体,将大麻哈鱼肝总DNA的15-20Kb片段重组到载体中,再包装成新的重组噬菌体,以含虹鳟鱼生长激素DNA部分序列的PAF51为探针,经[^(12)P]dCTP标记后,与噬菌斑杂交,自显影,获得3个阳性斑,复筛获得90％以上阳性斑,酶切回收生长激素基因片段,实验证明,已成功地获得大麻哈鱼生长激素基因片段,可供进一步亚克隆或转移研究使用。The paper is presented that a Salmon genomic DNA library was constructed and Salmon GH genes were cloned. The genomic DNA was isolated from SaJmon liver and digested partially by Sau3A. A range of 15-20kb DNA fragments were recovered and ligated with arms isolated from EMBL3 by EcoRI and BamHI. A total 1. 9 × 10 pfu/ug of DNA library was screened by[22P]-dCTP labeled probe, a 200bp rainbow trout cDNA from pAF51. Three positive plaques are isolated and there are 90% positive by rescreening. A 6. 5kb DNA fragment containing Salmon GH DNA was isolated and cloned for further study of subclone and gene expression.

关 键 词：大麻哈鱼 基因文库 基因克隆 

分 类 号：S188[农业科学—农业基础科学]