Construction Of Leaf And Fruit CDNA Library Of Lycium Barbarumand Isolation Of LycB And IPI Genes

Total RNA of leaf and fruit Lycium Barbarum L. was isolated using guanidinium isothioccyanate and phenol hloroform. mRNA was prepared by PolyATract mRNA Isolation Systems (Promega). Full條ength cDNA was synthesized by Universal RiboClone cDNA Synthesis System(Promega) and then cloned into phage # Excell vector, After Pachage in Packagene lambda DNA Packaging System, the capacities of the libraries were measured. The capacities of the libraries were 2.78 X 105pfu/ml and 8.4 X 104 pfu/ml. the recombination rates reached to 88.1% and 86.9%.Heterologous probe of Lye-. IPI PSY, ZDS, LycE originate from Getina lutea were labeled with digoxingenin-11-dUTP (DIG), were further used to screen the cDNA libraries of leaf and fruit of lycium barbarm. Eiight lycB positive clones were obtained and seven IPI positive clones were obtained. The recombinants with the cDNA insert fragment were released from the positive clones and were digested with EcoR I , The cDNA fragment were 2.1kb and 1.5kb in lycB and IPI positive clones respectively. Thisstudy will lay foundations for regulating carotenoid biosynthesis via genetic engineening.