中国生物工程杂志

In order to study gene express related to flower development of Lycoris longituba，total RNA from the petals of Lycoris longituba was extracted and purified by using the modified acid guanidinium thiocyanale-phenol-chloroform extraction method and the mRNA was isolated from total RNA by using Biotin-labeled Oligo(dT) and SAPMPs. The cDNA was synthesized and catalysed by reverse transcripase. The cDNA was then linked with EcoRⅠAdaptor. And the ends were phosphorylated. The Xho Ⅰdigestionreleased EcoRⅠAdaptor. The fragments smaller than 500 bp were removed by gel-purification and ligated to vector to transform the host by electroporator。 Finally the cDNA library of Lycoris longituba was constructed. The sink size of the primary cDNA is 1.112×106, in which 96.8% phages were recombinant, insert sizes ranging from 0.8～3.0kb were obtained. All of the above mentioned accorded with the general requirements of cDNA library construction, which established a good foundation for investigating the flower development of Lycoris longituba and cloning full length DNA of related genes with the methodology of both genomics and molecular biology.