虹鳟鱼(Salmo grairdneri Richard)基因文库的构建及生长激素基因的克隆和亚克隆  

Construction of Rainbow Trout Gene Library and Cloning and Subcloning of its Growth Hormone Gene

作　　者：杨学成[1] 杨志兴[1] 毕吉威 刘伟民[1] 赵晓祥[1] 王革伏 

机构地区：[1]黑龙江省科学院应用微生物研究所 [2]山东威海市医药公司

出　　处：《生物技术》1991年第6期10-14,共5页Biotechnology

基　　金：黑龙江省科学院资助

摘　　要：以噬菌体EMBL3作为载体,构建了虹鳟鱼的全基因文库,制备了包装蛋白效率达4.6×10~8Pfu/μgDNA.基因文库重组噬菌的量为8.2×10~5pfu/μgDNA.同时用含虹鳟鱼生长激素基因cDNA的pAF-51△S作为探针,从该文库中筛选出两个阳性噬斑,复筛获得90％以上的阳性斑。并从阳性克隆中回收虹鳟收生长激素基因片段为6kb,亚克隆到puc-18质粒中,进行了酶切分析。A library of rainbow trout genomic DNA was constructed by means of in vitro packaging procedure.Packaging efficiency of 8.2×05pfuof recombinated DNA have been achieved.The packaging protein with efficiency of 4.6×108pfu was preparated in this laborotory.A Plasmid PAF51ΔS containing 200bp rtGH sequence was used as a probe for on situ hybidization from 3 × 106 plaques of rainbow trout genomic library. Several positive plaques were screened and rescreened.The result shows that a gene containing rainbow trout growth hormone was cloned and subcloned. It can be used for further research work.

关 键 词：虹鳟鱼 基因文库 克隆 亚克隆 

分 类 号：S188[农业科学—农业基础科学]