光敏核不育籼稻 Hs-1 可育和不育幼穗 cDNA 文库的构建

The Construction of Fertile and Sterile Young Panicle cDNA Libraries of Photo sensitive Genic Male Sterile Rice Hs 1

分别取光敏核不育籼稻Ｈｓ－１在可育（１０ｈ／１７℃）和不育（１４ｈ／２７℃）条件下育性转换敏感期１—３ｃｍ幼穗为材料，以λＴｒｉｐｌＥｘ为载体，经离体包装构建成一对可育和不育ｃＤＮＡ文库，该文库具有构建快速、重组率高以及便于插入片段的亚克隆及其表达分析等特点。讨论了文库的应用前景，认为构建的两个文库用于筛选高丰度表达的光敏核不育基因是可行的。

The fertile and sterile total RNA and their mRNA were prepared respectively by Qiagen RNeasy kit from 1 3cm young panicles of photo sensitive genic male sterile rice Hs 1 in the photo period sensitive stage of fertility transformation in conditions of 10h/27℃ and 14h/27℃. Their full length cDNA were synthesized and then cloned into phage λTriplEx vector employing Clontech kit, USA. After packaged in vitro, the capacities of the libraries were measured. The capcity of fertile cDNA library was 2.2×10 5 pfu, and the recombination rate was 81.8%; The capacity of sterile cDNA library was 1.02×10 5 pfu and the recombination rate was also 81.8%. This paper also discusses the practical prospects of the libraries and points out that it is feasible to screen photoperiod sensitive genic male sterile genes expressed abundantly by using the libraries.