马铃薯基因文库的构建及贮藏蛋白基因的筛选

CONSTRUCTION OF POTATO(SOLONUME TUBEROSOME)GENOMIC LIBRARY AND SCREEN AND ISOLATION OF PATATIN GENE

以大容量的ＬｏｒｉｓｔＸ为载体，与马铃薯总ＤＮＡ部分酶切产物３０～５０Ｋｂ片段连接，经体外包装和效价测定，得到了６．５×１０５重组克隆，达到了构建马铃薯基因文库的要求．ＬｏｒｉｓｔＸ与ｐｕｃ９质粒可构成筛选目的基因的体内同源重组系统．利用这一系统，经筛选得到了４个目的基因克隆，经分析鉴定，４个克隆的插入片段均含目的基因或其片段．同时也表明这一同源重组系统在真核生物基因筛选方面是有效的．

The Potato genomic library was constructed with the Lorist X vector,which is a higher capacity vector in field of gene enginnering. By packaging in vivo and titering, 6. 5 × 10￣5recombinants had been got,and it contain about 99% DNA of whole Potato genome. The puc9 plasmid and Lorist X constitute a gene screen system of homologous recombination,by which,we got 4 clones. Our results showed that the Patatin gene or the gene fragments was inserted in each of these 4 clones,and this system may be efficient for gene screen from eucaryote genome library.